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Nanobiomaterials in
Soft Tissue
Engineering
Applications of
Nanobiomaterials
 Nanobiomaterials in
Soft Tissue
Engineering
Applications of
Nanobiomaterials

Edited by
Alexandru Mihai Grumezescu
Department of Science and Engineering of Oxide Materials and
Nanomaterials, Faculty of Applied Chemistry and Materials Science,
University Politehnica of Bucharest, Bucharest, Romania

Department of Biomaterials and Medical Devices, Faculty of Medical

Engineering, University Politehnica of Bucharest, Bucharest, Romania

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List of contributors

Gustavo A. Abraham
Research Institute for Materials Science and Technology, INTEMA (UNMdP-
CONICET), Mar del Plata, Argentina
Madalina Georgiana Albu
National Research & Development Institute for Textiles and Leather
(INCDTP) Leather and Footwear Research Institute, Bucharest, Romania
Ecaterina Andronescu
Department of Science and Engineering of Oxide Materials and Nanomaterials,
Faculty of Applied Chemistry and Materials Science, University Politehnica of
Bucharest, Bucharest, Romania
Florencia Montini Ballarin
Research Institute for Materials Science and Technology, INTEMA
(UNMdP-CONICET), Mar del Plata, Argentina
Vaishali Bambole
Department of Physics, University of Mumbai, Santacruz (E), Mumbai, India
Pierre Basmaji
Innovatec’s—Biotechnology Research and Development, São Carlos, SP, Brazil
Cornelia Bejenaru
Department of Vegetal & Animal Biology, Faculty of Pharmacy, University of
Medicine and Pharmacy of Craiova, Craiova, Romania
Ludovic Everard Bejenaru
Department of Pharmacognosy & Phytotherapy, Faculty of Pharmacy,
University of Medicine and Pharmacy of Craiova, Craiova, Romania
Şerban Vifor Gabriel Beresteanu
Head & Neck Surgery Clinic, Colţea Clinical Hospital, Bucharest, Romania
Pablo C. Caracciolo
Research Institute for Materials Science and Technology, INTEMA (UNMdP-
CONICET), Mar del Plata, Argentina
Michele Carrabba
Research Center E. Piaggio, University of Pisa, Pisa, Italy
Mariana Carmen Chifiriuc
Microbiology Immunology Department, Faculty of Biology, University of
Bucharest, Bucharest, Romania; Research Institute of the University of
Bucharest, Bucharest, Romania

xv
xvi List of contributors

Pablo R. Cortez Tornello

Research Institute for Materials Science and Technology, INTEMA (UNMdP-
CONICET), Mar del Plata, Argentina
Ligia Maria Manzine Costa
Department of Chemistry USP/FFCLRP, Brazil
Giuseppe Criscenti
Research Center E. Piaggio, University of Pisa, Pisa, Italy
Carmen Curutiu
Microbiology Immunology Department, Faculty of Biology, University of
Bucharest, Bucharest, Romania; Research Institute of the University of
Bucharest, Bucharest, Romania
Yaser Dahman
Department of Chemical Engineering, Ryerson University, Toronto, Canada
Gildásio de Cerqueira Daltro
College Hospital Complex Prof. Edgard Santos (COM-HUPES), UFBA,
Salvador, Brazil
Paula Braga Daltro
Department of Physical Chemistry UNESP/Araraquara-SP, Brazil
Aurora De Acutis
Research Center E. Piaggio, University of Pisa, Pisa, Italy
Carmelo De Maria
Research Center E. Piaggio, University of Pisa, Pisa, Italy; Department of
Ingegneria dell’Informazione, University of Pisa, Pisa, Italy
Lia-Mara Ditu
Microbiology Immunology Department, Faculty of Biology, University of
Bucharest, Bucharest, Romania; Research Institute of the University of
Bucharest, Bucharest, Romania
Filová Eva
Department of Tissue Engineering, Institute of Experimental Medicine of the
ASCR, v. v. i., Prague, Czech Republic
Prosecká Eva
Department of Tissue Engineering, Institute of Experimental Medicine of the
ASCR, v. v. i., Prague, Czech Republic
Anton Ficai
Department of Science and Engineering of Oxide Materials and Nanomaterials,
Faculty of Applied Chemistry and Materials Science, University Politehnica of
Bucharest, Bucharest, Romania
 List of contributors xvii

Denisa Ficai
Department of Inorganic Chemistry, Faculty of Applied Chemistry and
Materials Science, University Politehnica of Bucharest, Bucharest, Romania
Garazi Gainza
NanoBioCel Group, Laboratory of Pharmaceutics, University of the Basque
Country (UPV/EHU), School of Pharmacy, Vitoria, Spain; Biomedical Research
Networking Center in Bioengineering, Biomaterials and Nanomedicine (CIBER-
BBN), Vitoria, Spain
Itxaso Garcia-Orue
NanoBioCel Group, Laboratory of Pharmaceutics, University of the Basque
Country (UPV/EHU), School of Pharmacy, Vitoria, Spain; Biomedical Research
Networking Center in Bioengineering, Biomaterials and Nanomedicine
(CIBER-BBN), Vitoria, Spain
Maryam Ghaffari
Biomaterials Group, Faculty of Biomedical Engineering (Center of Excellence),
Amirkabir University of Technology, Tehran, Iran
Mustafa Abu Ghalia
Department of Chemical Engineering, Ryerson University, Toronto, Canada
Mazaher Gholipourmalekabadi
Biotechnology Department, School of Advanced Technologies in Medicine,
Shahid Beheshti University of Medical Sciences, Tehran, Iran
Aaron S. Goldstein
Department of Chemical Engineering, Virginia Tech, Blacksburg, VA, USA;
School of Biomedical Engineering and Sciences, Virginia Tech, Blacksburg, VA,
USA
Raluca Grigore
Head & Neck Surgery Clinic, Colţea Clinical Hospital, Bucharest, Romania
Alexandru Mihai Grumezescu
Department of Science and Engineering of Oxide Materials and Nanomaterials,
Faculty of Applied Chemistry and Materials Science, University Politehnica of
Bucharest, Bucharest, Romania
Antonio Carlos Guastaldi
Department of Physical Chemistry UNESP/Araraquara-SP, Brazil
Rosa Maria Hernandez
NanoBioCel Group, Laboratory of Pharmaceutics, University of the Basque
Country (UPV/EHU), School of Pharmacy, Vitoria, Spain; Biomedical Research
Networking Center in Bioengineering, Biomaterials and Nanomedicine (CIBER-
BBN), Vitoria, Spain
xviii List of contributors

Alina Maria Holban

Department of Science and Engineering of Oxide Materials and Nanomaterials,
Faculty of Applied Chemistry and Materials Science, University Politehnica of
Bucharest, Bucharest, Romania; Microbiology Immunology Department,
Faculty of Biology, University of Bucharest, Bucharest, Romania; Research
Institute of the University of Bucharest, Bucharest, Romania
Manoli Igartua
NanoBioCel Group, Laboratory of Pharmaceutics, University of the Basque
Country (UPV/EHU), School of Pharmacy, Vitoria, Spain; Biomedical Research
Networking Center in Bioengineering, Biomaterials and Nanomedicine (CIBER-
BBN), Vitoria, Spain
Carmen Iordache
Microbiology Immunology Department, Faculty of Biology, University of
Bucharest, Bucharest, Romania; Research Institute of the University of
Bucharest, Bucharest, Romania
Vahid Jajarmi
Biotechnology Department, School of Advanced Technologies in Medicine,
Shahid Beheshti University of Medical Sciences, Tehran, Iran; Cellular and
Molecular Biology Research Center, Shahid Beheshti University of Medical
Sciences, Tehran, Iran
Veronica Lazar
Microbiology Immunology Department, Faculty of Biology, University of
Bucharest, Bucharest, Romania; Research Institute of the University of
Bucharest, Bucharest, Romania
Plencner Martin
Department of Tissue Engineering, Institute of Experimental Medicine of the
ASCR, v. v. i., Prague, Czech Republic
Daniela Meliţă
Department of Science and Engineering of Oxide Materials and Nanomaterials,
University Politehnica of Bucharest, Bucharest, Romania
Rampichová Michaela
Department of Tissue Engineering, Institute of Experimental Medicine of the
ASCR, v. v. i., Prague, Czech Republic
Holeček Miroslav
New Technologies Research Centre, University of West Bohemia, Pilsen,
Czech Republic
 List of contributors xix

Laurenţiu Mogoantă
Research Center for Microscopic Morphology and Immunology, University of
Medicine and Pharmacy of Craiova, Craiova, Romania
George Dan Mogoşanu
Department of Pharmacognosy & Phytotherapy, Faculty of Pharmacy,
University of Medicine and Pharmacy of Craiova, Craiova, Romania
Masoud Mozafari
Bioengineering Research Group, Nanotechnology and Advanced Materials
Department, Materials and Energy Research Center (MERC), Tehran, Iran
Gabriel Molina de Olyveira
Department of Physical Chemistry UNESP/Araraquara-SP, Brazil
Satish Patel
University Institute of Pharmacy, Pt. Ravishankar Shukla University, Raipur,
Chhattisgarh, India
Jose Luis Pedraz
NanoBioCel Group, Laboratory of Pharmaceutics, University of the Basque
Country (UPV/EHU), School of Pharmacy, Vitoria, Spain; Biomedical Research
Networking Center in Bioengineering, Biomaterials and Nanomedicine (CIBER-
BBN), Vitoria, Spain
Kochová Petra
European Centre of Excellence NTIS, Faculty of Applied Sciences, University of
West Bohemia, Pilsen, Czech Re/public
Marcela Popa
Microbiology Immunology Department, Faculty of Biology, University of
Bucharest, Bucharest, Romania; Research Institute of the University of
Bucharest, Bucharest, Romania
Gabriela Purcel
Department of Science and Engineering of Oxide Materials and Nanomaterials,
University Politehnica of Bucharest, Bucharest, Romania
Zahra Rezvani
Bioengineering Research Group, Nanotechnology and Advanced Materials
Department, Materials and Energy Research Center (MERC), Tehran, Iran
Carla dos Santos Riccardi
Department of Physical Chemistry UNESP/Araraquara-SP, Brazil
Márcio Luiz dos Santos
Department of Physical Chemistry UNESP/Araraquara-SP, Brazil
xx List of contributors

Haji Shirinzadeh
Device Fabrication Group, Semiconductors Department, Materials and Energy
Research Center (MERC), Tehran, Iran
Deependra Singh
University Institute of Pharmacy, Pt. Ravishankar Shukla University, Raipur,
Chhattisgarh, India; NCNPR, School of Pharmacy, University of Mississippi,
Oxford, MS, USA
Manju Rawat Singh
University Institute of Pharmacy, Pt. Ravishankar Shukla University, Raipur,
Chhattisgarh, India
Maria Sonmez
University Politehnica of Bucharest, Bucharest, Romania; National Research &
Development Institute for Textiles and Leather (INCDTP) Leather and
Footwear Research Institute, Bucharest, Romania
Kubı́ková Tereza
Department of Histology and Embryology, Faculty of Medicine in Pilsen,
Charles University, Prague, Czech Republic
Patrick S. Thayer
School of Biomedical Engineering and Sciences, Virginia Tech, Blacksburg, VA,
USA
Keshav Deo Verma
Department of Physics, S.V. (P.G.) College, Aligarh, Uttar Pradesh, India
Silvia Villullas
Biopraxis Research AIE, Miñano, Spain
Giovanni Vozzi
Research Center E. Piaggio, University of Pisa, Pisa, Italy; Department of
Ingegneria dell’Informazione, University of Pisa, Pisa, Italy
Jatinder Vir Yakhmi
HomiBhabha National Institute (DAE), Mumbai, India
Tonar Zbyněk
Biomedical Centre, Faculty of Medicine in Pilsen, Charles University, Prague,
Czech Republic
Preface of the series

The era of nanosized materials is now considered the center of the evolution of
future tools and emerging technologies with wide applications in industry,
research, health, and beyond. Despite recent scientific progress, biological appli-
cations of nanomaterials are far from being depleted and current knowledge is
limited by the poor access to significant data, but also by widespread and usually
unfounded speculation. Although exhaustive, the current literature is difficult to
reach and understand because of the specificity and strict focuses of researchers
investigating different applications of nanomaterials.
In this context, the scientific series entitled “Applications of
Nanobiomaterials” was motivated by the desire of the Editor, Alexandru Mihai
Grumezescu, and others to bring together comprehensive, up-to-date and relevant
findings on the field of biological applications of nanostructured materials, to pro-
mote the knowledge and expand our vision regarding future perspectives. Even
though the approached domain is quite specific and research-oriented, this multi-
volume set is easily intelligible for a wide audience including: under-graduate and
post-graduate students, engineers, researchers, academic staff, pharmaceutical
companies, biomedical sector, and industrial biotechnologies. However, some
basic knowledge of the field of materials science (nanobiomaterials, pharmaceuti-
cal industry, products for medicinal treatments, nanoarchitectonics for delivery of
biological active molecules and release, bone implants and stomatology) and
engineering is a requisite for understanding technical aspects.
The selected authors of each chapter are outstanding specialists in the field of
nanobiomaterials, who have made impressive contributions in a specific area of
research or applied area within the scope of this book.
Each of the 11 volumes of the series contains 15 chapters, addressing the most
relevant and recent matters on the field of the volume.
The first volume, Fabrication and Self-Assembly of Nanobiomaterials, intro-
duces the reader to the amazing field of nanostructured materials and offers inter-
esting information regarding the fabrication and assembly of these nanosized
structures. In Volume II, entitled Engineering of Nanobiomaterials, readers can
easily find the most commonly investigated methods and approaches for obtaining
tailored nanomaterials for a particular application, especially those with a great
deal of significance in the biomedical field. In the following step, readers will dis-
cover the importance and the ways of modifying the surface of nanostructured
materials to obtain bioactive materials, by reading Volume III, Surface Chemistry
of Nanobiomaterials. Starting with Volume IV Nanobiomaterials in Hard Tissue
Engineering and Volume V Nanobiomaterials in Soft Tissue Engineering the bio-
medical applications of engineered nanomaterials are revealed and discussed,
focusing on one of the most impacted fields, tissue engineering. Volume VI,
Nanobiomaterials in Antimicrobial Therapy, highlights the potential of different

xxi
xxii Preface of the series

nanostructured materials to be utilized in the development of novel efficient anti-

microbial approaches to fight the global crisis of antibiotic inefficiency and
emerging infectious diseases caused by resistant pathogens. Volume VII moves
on to another key biomedical domain—cancer therapy. This volume,
Nanobiomaterials in Cancer Therapy, describes current issues of cancer therapy
and discusses the most relevant findings regarding the impact of nanobiomaterials
in cancer management. Medical Imaging represents the focus of Volume VIII,
while Volume IX deals with applications of Nanobiomaterials in Drug Delivery.
Volume X, entitled Nanobiomaterials in Galenic Formulations and Cosmetics,
refers to the perspectives highlighted by the utilization of nanosized functional
biomaterials in the development of improved drugs and active principles for dif-
ferent biomedical industries. Finally, Volume XI is dedicated to the impact of
Nanobiomaterials in Dentistry, which currently represents one of the most investi-
gated and controversial domains related to the biomedical applications of nanos-
tructured materials.
Due to their specific organization, each volume can be treated individually or
as a part of this comprehensive series, which aims to bring a significant contribu-
tion to the field of research and biomedical applications of nanosized engineered
materials.
Ecaterina Andronescu
Department of Science and Engineering
of Oxide Materials and Nanomaterials,
Faculty of Applied Chemistry and Materials Science,
University Politehnica of Bucharest, Bucharest, Romania
Preface

ABOUT THE SERIES (VOLUMES I XI)

The increased fabrication of nanosized materials with applications on the biomed-
ical field by using biomimetic and bioinspired processes and formulations has
recently led to a new concept, nanobiotechnology. This complex research brings
together significant knowledge from physical, chemical, biological, and techno-
logical sciences in an applicative field.
Medical applications of nanobiomaterials range from the development of ade-
quate scaffolds for tissue engineering to therapeutic nanostructures, such as tar-
geted drug-delivery systems. The purpose of this multivolume set entitled
Applications of NanoBioMaterials is to offer a broad, updated, and interdisciplin-
ary point of view regarding the applications of these “materials of the future med-
icine,” starting with their fabrication, specific engineering, and characterization
and ending with the most investigated applications such as tissue engineering,
antimicrobial and cancer therapies, and also the development of different medical
and cosmetic products. These books bring together the work of outstanding con-
tributors who have significantly enhanced the basic knowledge and applicative
concepts of this research field in their respective disciplines.
The multivolume set Applications of NanoBioMaterials contains 165 chapters,
organized in 11 volumes which are ready to present a novel and up-to-date
approach related to this intriguing domain. Each chapter is carefully composed
and illustrated to highlight the relevance of nanobiomaterials on relevant biomedi-
cal fields, revealing the most recent applications on each investigated domain.
The whole set represents a great material for the academic community, starting
with undergraduate and postgraduate students, researchers, engineers, and medical
doctors, but also pharmaceutical companies, the industrial sector, and innovative
biotechnologies.
These 11 volumes cover almost all aspects related to the Applications of
NanoBioMaterials as it follows:
Volume I: Fabrication and Self-Assembly of Nanobiomaterials
Volume II: Engineering of Nanobiomaterials
Volume III: Surface Chemistry of Nanobiomaterials
Volume IV: Nanobiomaterials in Hard Tissue Engineering
Volume V: Nanobiomaterials in Soft Tissue Engineering
Volume VI: Nanobiomaterials in Antimicrobial Therapy
Volume VII: Nanobiomaterials in Cancer Therapy
Volume VIII: Nanobiomaterials in Medical Imaging
Volume IX: Nanobiomaterials in Drug Delivery
Volume X: Nanobiomaterials in Galenic Formulations and Cosmetics
Volume XI: Nanobiomaterials in Dentistry

xxiii
xxiv Preface

ABOUT VOLUME V
Volume V, NanoBioMaterials in Soft Tissue Engineering, is an up-to-date book
that presents the recent progress of dedicated nanobiostructures in soft tissue engi-
neering. The book highlights the main conventional and unconventional fabrication
methods of typical three-dimensional (3D) scaffolds used in regenerative medicine
(such as for muscle, nerve, tendon, ligament disorders, cartilage, neural outgrowth
and regeneration, cardiovascular diseases, skin burns, and wounds). Also, in this
work is presented an up-to-date overview regarding surface modification, spatial
properties, and the newest in vivo applications of engineered 3D scaffolds.
Volume V contains 15 chapters, prepared by outstanding international
researchers from the United States of America, Argentina, Brazil, Spain, Italy,
Czech Republic, Romania, Iran, and India.
In Chapter 1, Soft Tissue Engineering and Microbial Infections: Challenges
and Perspectives, Carmen Mariana Chifiriuc et al. review the recent progress in
tissue engineering applications from the design of multifunctional biomaterials
assuring simultaneously the scaffold function, as well as the release of antimicro-
bial drugs, growth factors, and other bioactive molecules, in order to prevent
infections and therefore to accelerate optimal tissue regeneration.
Garcia-Orue Itxaso et al., in Chapter 2, Nanotechnological Approaches for
Skin Wound Regeneration Using Drug-Delivery Systems, discuss engineered nano-
materials utilized in delivery and controlled release of drugs used in skin wound
regeneration, particularly emphasizing with polymeric and also lipid nanoparticles,
silver nanoparticles, nanofibrous structures, nanosheets, and nanohybrids.
Chapter 3, Bacterial Cellulose for Advanced Medical Materials, prepared by
Gabriel Molina de Olyveira et al. describes recent advances regarding the applica-
tions of bacterial cellulose in human regenerative medicine and stem cell behav-
ior. The chapter also reveals recent drug-delivery applications and future insights
with respect to bacterial cellulose.
In Chapter 4, Applications of Nanobiopolymers for Soft Tissue Engineering,
George Dan Mogoşanu et al. present details about the main natural and synthetic
polymers used for soft tissue engineering: collagen, chitosan, heparin, fibrin,
gelatin, alginate, hyaluronic acid, silk; polylactic acid, polyglycolic acid, poly
(lactic-co-glycolic acid), poly-ε-caprolactone, poly(L-lactic acid-co-ε-caprolac-
tone), polyethylene oxide, polyurethane, polyethylene terephthalate, and
polymethyl methacrylate, as well as their clinical applications.
Next, C. De Maria et al., in Chapter 5, Machine Design for Multimaterial
Processing, present the design of a novel parallel manipulator with three transla-
tional degrees of freedom for microfabrication of biomaterials. In this work,
hydrogel-based composite scaffolds with applications in soft tissue engineering
are discussed.
In Chapter 6, Advanced Nanobiomaterials in Tissue Engineering: Synthesis,
Properties, and Applications, Mustafa Abu Ghalia and Yaser Dahman highlight
 Preface xxv

the variety of organic and inorganic materials used in tissue engineering

applications. Fabrication routes and details of the processing techniques used to
evaluate interfaces in tissue engineering are also provided.
Gabriela Purcel et al., in Chapter 7, Collagen-Based Nanobiomaterials:
Challenges in Soft Tissue Engineering, describe the main properties, advantages,
and biomedical applications of collagen. The chapter highlights functional nano-
biomaterials based on collagen for use in regenerative medicine, not only as a
subject for laboratory studies, but also for general clinical use.
In Chapter 8, Micro/Nanofiber-Based Scaffolds for Soft Tissue Engineering
Applications: Potential and Current Challenges, Pablo R. Cortez Tornello et al.
give an overview about recent research advances made in the development of
electrospun scaffolds for soft tissue engineering applications focusing on wound
dressings, cartilage, muscle, cardiovascular, nerve, and skin tissues.
Chapter 9, Natural Polymer-Based Hydrogels as Scaffolds for Tissue
Engineering, prepared by Manju Rawat Singh et al., focuses on fabrication and
applications of hydrogel scaffolds obtained with natural polymers used in the treat-
ment of wounds and healing, with the ultimate goal of clinical tissue regeneration.
Chapter 10, Bioactive Nanomaterials for Cartilage and Muscle Regeneration,
prepared by Alina Maria Holban et al., presents the most prevalent cartilage- and
muscular tissue-related diseases and highlights the recent scientific progress
regarding the development of classical and recent therapeutic approaches, also
discussing the impact and advantages of bioactive nanomaterials in designing
functional scaffolds for cartilage and muscle repair.
Aaron S. Goldstein and Patrick S. Thayer, in Chapter 11, Fabrication of
Complex Biomaterial Scaffolds for Soft Tissue Engineering by Electrospinning,
present an up-to-date overview about the electrospinning process for the
engineering of specific soft tissues, including muscle, nerve, tendon, ligament,
skin, and vascular applications. This chapter reviews fabrication methods of micro-
fiber meshes through electrospinning and describe how changing the fiber properties
can influence cell behavior through contact guidance and mechanical effects.
Chapter 12, Oxygen-Generating Nanobiomaterials for the Treatment of
Diabetes: A Tissue Engineering Approach, by M. Gholipourmalekabadi et al.,
discusses the impact of oxygen-generating nanobiomaterials in the improvement
of survival of transplanted cellular products by sustained oxygen delivery within
the microenvironment of tissue-engineered sites.
Denisa Ficai et al., in Chapter 13, Advances in the Field of Soft Tissue
Engineering: From Pure Regenerative to Integrative Solutions, report the recent
progress on regenerative systems successfully combined with therapeutic agents
in order to design synthetic graft materials used in the treatment of skin infections
of skin cancer.
In Chapter 14, Tissue Engineering: Use of Electrospinning Technique for
Recreating Physiological Functions, Vaishali Bambole and J. V. Yakhmi discuss
aspects regarding tissue engineering in the electrospinning process and review the
most relevant and recent typical case studies.
xxvi Preface

In Chapter 15, In Vivo Microscopic and Mechanical Characteristics of

Bioengineered and Biodegradable Tissue Scaffolds and Nanomaterials, Kubı́ková
Tereza et al. present an up-to-date overview of chemical composition, dimensions,
surface modifications, and spatial properties (spacing and porosity) of meshes and
fiber scaffolds. The authors discuss the modifications of scaffolds that affect
adhesion, migration, proliferation, and differentiation of cells as well as formation
of new extracellular matrix.
Alexandru Mihai Grumezescu
Department of Science and Engineering
of Oxide Materials and Nanomaterials,
Faculty of Applied Chemistry and Materials Science,
University Politehnica of Bucharest,
Bucharest, Romania

Department of Biomaterials and Medical Devices,

Faculty of Medical Engineering,
University Politehnica of Bucharest, Romania
http://grumezescu.com/
 CHAPTER

Soft tissue engineering and

microbial infections:
challenges and perspectives 1
Mariana Carmen Chifiriuc1, Anton Ficai2, Alexandru Mihai Grumezescu2,
Lia-Mara Ditu1,4, Marcela Popa1, Carmen Iordache1, Alina Maria Holban1,2,4,
Şerban Vifor Gabriel Beresteanu3, Raluca Grigore3 and Veronica Lazar1
1
Microbiology Immunology Department, Faculty of Biology, University of Bucharest,
Bucharest, Romania 2Department of Science and Engineering of Oxide Materials and
Nanomaterials, Faculty of Applied Chemistry and Materials Science, University Politehnica of
Bucharest, Bucharest, Romania 3Head & Neck Surgery Clinic, Colţea Clinical Hospital,
Bucharest, Romania 4Research Institute of the University of Bucharest, Bucharest, Romania

1.1 INTRODUCTION
Tissue engineering requires an interdisciplinary approach for the achievement of
successful strategies dedicated to human tissue reconstruction, regeneration, and
restoration of their normal functions in conditions in which their self-repairing
capacity is affected by disease or injury (Herbert and Rahul, 2005).
The identified practical solutions require the use of biomaterials as scaffolds
and/or delivery systems for cells, growth factors, and other bioactive molecules
stimulating the cellular proliferation, differentiation, and integration of new tissue.
The term “tissue engineering” refers to bone and cartilage or soft tissue regenera-
tion. This chapter is dedicated to the last of these two categories.
Tissue engineering could represent a solution by the construction of biological
substitutes which could be used instead of donated organs (Atala, 2004).
Soft tissue replacement needs can arise for various reasons, such as congenital
or chronic diseases (e.g., infections, inflammatory diseases, cystic fibrosis, ciliary
dyskinesia, cardiovascular, congenital or acquired diseases), tumor rejection or
other traumatic lesions (e.g., corneal scars, severe burns, and deep wounds), and
the clinical approach consists more often of autologous tissue grafts, and less fre-
quently in the use of allografts, xenografts, and synthetic materials, in order to
avoid undesired complications, such as infection or graft rejection (Stosich et al.,
2009).
Some of the limits of conventional approaches are expected to be resolved by
tissue engineering technologies, although this will not be an easy task to achieve,
Nanobiomaterials in Soft Tissue Engineering. DOI: http://dx.doi.org/10.1016/B978-0-323-42865-1.00001-5
© 2016 Elsevier Inc. All rights reserved.
1
 1.4 Antimicrobial Polymers Used in Soft Tissue Engineering 13

occurrence of a corneal rejection reaction, neovascularization, or keratoconus in

the following 3 months (Liu et al., 2014).
An infection-preventing membrane composed of a dense and porous collagen
network has been developed. A bilayered membrane composed of hyaluronan
microparticles loaded with silver-sulfadiazine was fabricated by gelling and was
then incorporated into collagen layers. The collagen membrane assured a sus-
tained release of the antimicrobial agent in active form, as revealed by its inhibi-
tory activity against P. aeruginosa sustained for 4 days, while in vivo, the use of
such membrane was associated with increased tissue regeneration (Lee et al.,
2002).
A dual-functional composite with anticoagulant and antibacterial properties
based on heparinized silk fibroin and chitosan was synthesized showing antimi-
crobial activity against S. aureus, proving its potential to be used as a composite
biomaterial for blood contact devices (Wang et al., 2011a,b).
Lactoferrin is a bioactive globular protein with antimicrobial activity but also
with potent antiapoptotic and osteogenic activity mediated by the Wnt5a/PKA
pathway and the stabilization of β-catenin by rhLF dependent on the PKA/LRP6
signaling pathway. Recombinant human lactoferrin gels have been shown to sup-
port MC3T3 osteoblast cell viability, proliferation, and differentiation, as well as
phosphorylation of signaling proteins (Amini and Nair, 2014).
Poly(ε-caprolactone) and gelatin blended with metronidazole designed for
guided tissue regeneration revealed a good release of the antibiotic, which suc-
cessfully prevented colonization with anaerobic bacteria (Xue et al., 2014).
Fibrous structures and synthetic polymer blends are promising candidates for
variate tissue-engineering applications (scaffolds for stem cells, engineered grafts,
vascular stents, drug delivery systems, transdermal patches, wounds, and burn
care).
The polyvinyl alcohol combination with chitosan also exhibited a large spec-
trum of antibacterial activities, thus protecting against infection (Bhullar et al.,
2015).
A bionanofilm consisting of homogeneous silver nanoparticles coated on
single-walled carbon nanotubes, which were later hybridized with DNA and stabi-
lized in poly(vinyl alcohol), proved to be biocompatible and displayed antimicro-
bial activity (Subbiah et al., 2011).
Bioactive adhesive materials containing dimethylaminododecyl methacrylate
exhibited a marked antibiofilm activity against Streptococcus mutans,
Streptococcus gordonii, and Streptococcus sanguinis biofilms (Zhang et al.,
2015).
The pH-sensitive, negatively charged layer-by-layer hydrogel films, consisting
of poly(2-alkylacrylic acids) crosslinked with varying polymethacrylic acid, poly
(2-ethylacrylic acid), poly(2-n-propylacrylic acid), and poly(2-n-butylacrylic
acid), have been proved to turn from hydrophilic to hydrophobic, becoming dehy-
drated and thus bactericidal in response to acidic conditions induced by bacterial
multiplication. The films were also highly biocompatible, proving to be promising
14 CHAPTER 1 Soft tissue engineering and microbial infections

candidates for the development of biomaterials with low infection risk (Lu et al.,
2015).
Biocompatible propolis-loaded polyurethane nanofibers exhibited significant
antibacterial activity, revealing a great potential for wound dressing and skin tis-
sue engineering (Kim et al., 2014).
Citrate-based polymers have unique properties recommending them for bio-
medical applications: citric acid is a natural biocompatible product with antimi-
crobial activity and also provides multiple functional groups for crosslinking and
for conjugation of biomolecule antimicrobial properties. Citric acid polymers sig-
nificantly reduced S. aureus and E. coli proliferation, this activity representing a
great advantage of this type of polymer for use in tissue engineering (Su et al.,
2014).
It has been shown that doxycycline antibiotic treatment for 1 month after her-
nia repair reduced the matrix metalloproteases and increased the collagen I:colla-
gen III ratios, and the tensile strength of repaired interface fascia (Tharappel
et al., 2014).
An antiadhesive polymer peptide conjugate composed of block copolymer
Pluronic F-127 and functionalized with antimicrobial peptides exhibited a contact
bactericidal effect against S. aureus, S. epidermidis, and P. aeruginosa, while the
arginine glycine aspartate peptides promoted the adhesion and spreading of
human fibroblast cells (Muszanska et al., 2014).
In situ forming hydrogels containing poloxamer grafted on the hyaluronic acid
were designed for tissue-engineering applications (i.e., ophthalmic delivery sys-
tem) and proved to assure a sustained release of ciprofloxacin (Cho et al., 2003).
Silver-nanoparticle-loaded nanofibrous PLGA scaffolds were obtained by
electrospinning and showed good biocompatibility and antimicrobial and antibac-
terial activity against S. aureus and K. pneumoniae at silver nanoparticle concen-
trations .0.5 wt%, proving their potential for tissue-engineering applications
(Xing et al., 2011).
Usnic acid-loaded PLGA-PVA-loaded microspheres proved to inhibit both the
initial S. aureus adherence phase as well as the development of mature biofilm
rendering them as competitive candidates for the development of scaffolds for
stem cell cultivation and tissue engineering (Grumezescu et al., 2014).
A nanostructured PLGA membrane designed to be used as a next-generation
antibacterial skin graft material proved to exhibit an intrinsic anti-S. aureus activ-
ity and, at the same time, promoted keratinocyte proliferation, without increasing
the density of dermal fibroblasts implicated in scar tissue formation, proving its
effectiveness in limiting scar tissue formation and bacterial invasion, while pro-
moting wound-healing (Karahaliloğlu et al., 2014).
Poly(D,L-lactide-co-glycolide) porous microspheres containing silver-releasing
degradable phosphate glass or metronidazole proved to exhibit antimicrobial
properties against S. aureus, E. coli, and Bacteroides fragilis, which normally col-
onize the perianal fistulae. Moreover, microspheres became rapidly integrated
within the host tissue after subcutaneous implantation into a rodent wound model,
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 —Vast woar,.. snapperden en pieperden de anderen
er door heen.

Er klonk wat dof gemor in ’t donker. Rink de reuzige

lobbes wou wèl. Willem ook en Jan ’t gulst. Maar Dirk
en Piet mokten koppig. Piet wou eerst nog bakkeleien.
—Dirk jeukten de handen. Ze gromden de twee
broers, als buldoggen. Plots in ’n ruk, drong Rink ze bij
elkaar, de stugge haatdragende wraakgierige neven.
En in lol plots aangekitteld door de lachschelle meisjes
sloeg hun haat in dronkemanszwaai over in zang,
werd hun versmoorde jaloerschheid in verteederden
gevoelszwenk, afgunstige vriendschap.—Willem
greep Dirk vast, Piet, Jan, Rink, de reuzige lobbes,
omarmde ’t heele stel, en Henk, ’n takknoest in de
hand, smakte zich brutaal met z’n donk’ren kop
tusschen de meiden, slobberde in ’n ommezientje
[285]ze de vroolijke wangen vol met klink-zoete
zoenen.

Jan Grint die stommetje had gespeeld strompelde op

zij, en met z’n afgeknakten boomtak, slingerde ie vóór
den stoet uit, tamboermajorig manoeuvreerend.

In slinger en sliert eindelijk joolden ze den weg af, op

stillen havenkant áán. De meiden vonden dat de
knapen zich kranig gehouden hadden. In joligend-
mekaar-aangekijk, met de vroolijke oogen vol lach en
rumoerlust, bonsden en hotsten ze den krommigen
keiweg af, schreeuwden ze ’t uit, toen ze plots vóór
zich den rooden schemer, nòg dichter bij den
brandgloed van al ’t gas, toortsgewalm en
kraamlampen zagen opvlammen. In genotssidder
hoorden ze klankenorkaan van al de orgels loeien en
dreunen, tusschen het menschkermend en scheurend
gezang van kermisgangers, die dromden rond den
kook van ’t avondlicht, den roffel van trommels, den
gil-schater van den stoomdraaimolen.

Van den polderweg àf, waar de luidlooze weien in

angstige luistering trilden en de stilte staàrde naar ’t
hellekrochtige kermisgerucht en fakkelgevlam, gierde
de stoet, hossend uit ’t zomerlijke duister, al dichter op
de licht-razernij áán. De meiden joolden, sprongen, de
kerels brulden.

Oooauuw! wat ’n ska-ànde

Loage wroak van En-gè-land!

Van de Haven klonk terug, zwak-vergalmend tot

stilstaand gerucht, naar ’t polderland: Ooaauw.. wat ’n
skande.—Uit alle havenhoeken daverde en galmde ’t
refrein áán; in stikdonker, bij schaduwrood schijnsel
van wat eenzame lantaarns; onder bronsgloed van
walmende toortsen. En van overal doemden òp in ’t
licht, zingende rooie koppen, schal-monden,
aangegloeid ros-oranjig in den hellen brand van
kermiswoel.—

—Binne dur nou puur luchtskommels? gulzigde Griet,

smorend ’r dolle blijheid nou ’t kermisgetier al dichter
op ’r aandruischen kwam, oorscheurend fel.—

Ze voelden zich dronken de meiden, nu al, van

koortsig verlangend [286]genot. Huiverwellust
schroeide ze’t lijf; genot kittelde ze in de kaken en al
dichter naderden ze ’t vulkanisch getoorts van gas en
 flonker-lampen, flambouwen en kleurig-schichtig
elektriek.

Sterker bulderden aan, orgeldreunen en ’n woeste

warrel en stuif van heete stemmenzang, krijsch en
fluitgegil spoog en hoosde ze in d’r begeertronies.

—Dur is dur t’met van alles Geert.… daa’s d’r puur

een prêcht! schreeuwde Willem opgewonden, en lievig
tegen z’n neef,—nie Dirk? dá’ hai jullie d’r vast nooit
sien.… dur is dur.. ’n reusin.… saife-honderd
pondjes.… en ’n ieder mog dur betaste en befoele.…
of dá’ sai dur ècht is.

—Jai weut d’r gain snars van, stampte Jan Grint

ertusschen,—je hep d’r ’n stoomdroaimole.. soo groot
krek aa’s ’n poardespul.… en je hep d’r ’n kie-me-tè-
mès-koop.… of hoe da loeder hiete mag.… je hep d’r
’n poardekirsis.… ’n skouburg en vaif kèfèe-setans.…
en ’n klodder goocheloars.. en woaresaisters.… en
skiettinte.. enne gruufelkoamers.. enne.…

—Hohee! gilde in schellen lach Rink er doorheen,

woar blaife de lekkere maide dan? wa kan main die
spullemikmak hoàmere.… De màide.… de maide.…
daa’s de klus! de fles.. dàas ’t faine werk.. aêrs f’rfail je
je aige stierlik!

—Ooaauuw! waa’t ’n ska-ande..

brulde Dirk midden in, met Willem die ’n maat

achterna strompelde; en hoog lolden de meiden mee.
—Hou d’r je snoater, nie veur je tait.. hakkekruk.. op
da terrain wee’k de sangbeweging te motte.. ikke
konsteteer van dâ ikke allainig de paàs sel merkeere
op dâ terrain.… hee Platneus.. hiere langst.… hiere
langst!.… Bolkie! En nou valle jullie d’r in.…

In bas-diepe brom zette Klaas Koome in.

’t Is de kerremis die je vreugte biet..

Van wa wai d’r nou rais lekker geniet! [287]
Wel sait d’r ’n droo-ooge—soo’n saa ie.. held’
Skaft d’r moar òf.. kost mooi je liefe geld.
Daa’t is d’r ’n la-amstroal van de ofskoàffersbond
’n Blaufe knoop in se jaa’s, gloasje mellik an sain
mond.

In stijgenden heeschen krijsch stond ie voor ’t eind en

heel de stoet sliertte ’t refrein de roodbegloeide lucht
in:

—Men singt, men host, men lacht, men moakt pelsier..

Je loa je waige of drink ’n gloasie bier!
Je soekt ’n maideke, da je spoedig vint!
In sukke doage binne sai goed gesind!
Nou goan je skiete in ’n linnen tint.
En roak ie waà’t, kraig je ’n mooi pirsint.

—Si-iint! galmden ze uit, meiden en kerels, vlak bij ’t

havenend waar ’n kankaneerende stoet uit
flambouwbrons weghoste, hen kruiste bij ’t duister
pakhuisgedrang, en rauw Klaas’ lied overschreeuwde
met ’t helsche refrein

—Oooaauw! waa’t ’n skande..

 Loage wroak van En-ge-land.

Loslatend hun eigen lied galmden de Grints en

Hassels mee met den rauwen stemmenzwenk der o-
wat-’n-skande-krijschers.—Zoo, dansend en wulpsch
schaterend hosten ze áán in duivelsch gebaar onder
eersten ros-gloed van hel gloed-verruischende
flambouwenrij, voor groote tent-façade.—

Besefloos werden ze plots oversmakt door ’n

bulderstroom van hossers. Met moeite werkten ze zich
weer bijéén uit den bruisenden golfstoot van zwarte,
duistere, èven en half-begloeide stoeten.

Vlak voor de luchtschommels, dicht aaneengehaakt in

armenknel, bleven ze eindelijk staan.

Een rij flakker-felle gasflambouwen op koperen

schitterstang gloeiden voor de schommeltent, in hel
van rood-goud licht verzwommen. Er naast, vóór de
kinematograaf boogden drie elektrische lampen, ’t
boomlommer van kastanjelaan ingeheschen,
[288]tusschen twijgen en zwaar geblader, rood-paarse
trilgloed verflakkerend, tooverig-dekoratief
rondlichtend òver den kermiswoel. Al lichtsoorten
vochten en worstelden tegen elkaar in. Aan één kant
vergloeiden de elektriekbollen als paarse manen, een
hel-klare prachtnevel, over boomen en menschkoppen
en tenten; aan anderen kant, vlak er naast, dromden
en gierden tronies, rossig-aangegloeid in beefwalm
van gasflambouwen en petroleumlampetten,
hellemaskers gekerfd en duivelsch geschminkt in ’t
licht en de angst-zware schaduwen; satanische
 storting van gloed op monden en oogen, kaken en
wangbrokken.—

Uit de nauwe steegjes worstelden lòs gistende

proppen bijeengeknelde kerels, meiden, jochies en
wijven. Achter de steegspleten met hun duister gewoel
gloedschijnselde brandroode lucht van Baanwijk, waar
vóórbrok van kermis joelde in helle lichtschatering.
Telkens nieuwe donkre drommen in de kronkelige
duistere gangetjes, stortten zich in den dampenden
avondgloei van vlam-spelige tenten en spullen. Lol,
zang en gebral botsten in rauwe hette onder den
elkaar-kruisenden menschenwoel in de donk’re
steegjes òp; gangetjes en kronkelspleten die er
duisterden als zwarte weggetjes tusschen twee rijken
vulkanisch beflambouwd: Baanwijk en Haven. Aan
ingang van wat kronkelgangetjes bloedde hier en daar
rood en groengeel gevlek, weerschijn van kleurig
tentglas op verweerde muurbrokken rond gekranst.
Maar de meesten donkerden als grotten, waar telkens
kankaneerende massa, uit ’t hellelicht van Baanwijk in
verzwelgde, voortwoelde ’n poos, in de grottennacht,
daar dobberde, terug bonkerde en weer voortgolfde;
gloeierig opdook plots weer in den fellen havenbrand,
den woest oranje-rood en paars-groenen gloed van
flonkerlampen, toortsen en elektriekbollen bij
kinematograaf, waarvoor ’t volk woelde klaar-belicht,
als in ’n plots opengebarsten dag.

Een orgelschallende orgie, demonische klankenroffel

en schetter tegen elkaar inzwirrelend, raasde rond in
hellesfeer op de Haven, kretenzee en brallende
geruchten, opgejaagd in de [289]gloeiheete tjink-tjinks
 van bekkenslag en pauken, lawaaiend tot àchter den
spoordijk waar polderland eenzaamde in eindeloozen
weischemer. En rondom, de melankolieke zeur van
ééndeunige orgelwijs. De kastanjeboomen, diepe laan
van schel-brandend groen, doorblauwd en doorpaarst
van fel-elektriek, grilligde daar als tooneelpark in
dekoratieven brand, met z’n dampig poortperspektief,
waar aan ’t eind, op verren achtergrond van
petroleumfakkelend oranjig roodgoud, een dolle dans
van demonen raasde, uitzinnige wereld van rood-
donk’re en rood-lichtende, kangoeroesch-
kankaneerende wezens; vrouwen en meiden
opschuimend d’r rokken in witte branding en bruising
van ondergoed; mannen en dronken kerels in vetten
lach of strammen ernst, kwijlend, verhit en omgloeid
met de klankrazernij van orgelschetter, paukendreun
en krijschzang.

Vóór de luchtschommels propten de meiden en kerels

zich òp, kanaljeus en doorschroeid van zinnepassie, in
puffende omarmingen, wachtend tot de klingelende
bel, ronde van elken luchtgang afrinkelde, de
roffelende trommel van matroosje vóór ’t orgel, plots
dòfte en de kas-juffer àfluien liet tusschen geknars van
de plots geremde schommels, gepiep van de stangen,
en den rumoerig-beenschuifelenden terugsmak der
stil-staande lijven uit den geweldigen slingergang.

In overrompelenden drom sprongen Dirk, Piet, Willem

en Rink met de meiden naar voren, fel-loerend op
plaats. Dirk had zich woest op Geerts slank lijf
gesmakt. Piet schommelde met schalksch-vurige Trijn,
de aanhalige Trijn, en Willem, woedend dat hij Geert
 niet handig genoeg mee had gesleurd, toch zich
goedhoudend, bonkerde er maar op los met de
snibbige Annie.—

Jan Grint had rondgekoekeloerd of ie Guurt ook zien

kon, al voelde ie vooruit, dat ze wel te trotsch zou zijn
om met den stoet mee te lollen.—

Rink had nog bij tijds ’n schommel gegrepen waarin ie

blonde zenuwachtige Cor droeg. Meer plaats was er
voor hun stoetje niet. Woedend, vol spijt holden de
overblijvenden terug van [290]de trap.—Klaas Koome
raasde wat tegen de luchtschippers die de sloepen
òpschoten, zonder den nijdas te woord te staan. Jan
Grint, Henk Hassel en de anderen dwarrelden
tusschen de aanhossende groepjes, toch niet te vèr
van hun klub, tukkig loerend op alleen staande
meiden, die meegesleurd wilden worden.

Bolk was verschrompeld en bleuïg, met z’n platten

polypneus berossigd en begroend, z’n grillig bevlamde
tronie de hoogte inkijkend, tusschen ’n troep meiden
weggeschuchterd; voelde zich niet meer thuis in ’t
lawaai. Dat hadden de kerels wel begrepen en
daarom ’m half aan zijn lot overgelaten. Op weg naar
de Grints hadden z’m ontmoet. Hij moest mee, of ie
wou of niet.

In ’n hos-storm was ie toen meegesleurd omdat ze

nog wat jool met den ouen rakker wilden hebben.—
Maar onder den wandel naar meiden en kermis wouên
ze’m al weer kwijt. Nou stond ie daar nog voor de
luchtschommel, bang-opkijkend naar de sloepjeszwier
hoog in de lucht,—in zijn tijd nooit bestaan—z’n ouden
 kop omkolkt van geraas, bemokerd van
dreungeluiden. Telkens schrok ie òp, dacht ie dat ’n
sloepje, rakelings langs ’m scherend, tegen z’n
harsens zou aanbonsen. En telkens weer verschool ie
z’n klein krom lijf schuchterder achter ’n woest-
gillenden meidenstoet, bijzij de tent verwoelend, rossig
aangegloeid in flakkering van flambouwenvlammen.

Dirk met Geert, ’t mooie zwartje, slingerde ’t hoogst

boven tien sloepen uit. Rink rukte als ’n bezetene aan
de stangen, dat ie plots in ontzettenden zwier-zwaai
met Cor boven tentdak uitzweefde.—In luchtzuigende
vaart schoten de schommels òp en néér, scheerden
den vloer, zwierden weer òp, plonsden weer neer uit
bang-duizelenden slingergang. En hooger, in woesten
naijver op elkaar, schreeuwend en lallend, zetten de
kerels òp, lijfzwaar zich schurend op de meidlijven bij
elken terugzwaai en smak naar àchter, de tent in.—

—Set d’r op Dirk! set d’r op! barstte Geert uit, in

heeten schater om haar hoog-dolle vaart, met rillige
gloeiingen in ’r lijf van schommelgenot, d’r wellust-
woeste oogen strak op Hassels koeienkop gebrand.
Dirk voelde ’t warme zweetlichaam [291]op zich
aanzwellen bij elken nieuwen ruk en inbuig van ’r
opzet. En rukken nu deed ie als ’n bezetene, dat de
stangen kermden in de scharnieren. ’n Woeste
zweeflol, ’n dolle zwier, niet voelend meer waar ie
was, golfde door Dirk’s handen en beenen; bloed
spoot ’m naar den kop. En Geert genoot mee, in
schommelzwijmel, bang-heerlijk, hoog-wèg in heeten
suizenden duizel, dàn in de lucht, dàn beneden,
scherend den planken vloer, dat al meer gloeiende
 rillingen door ’r heensidderden. Al de meiden in de
bootjes hitsten de kerels òp, hijgend, hooger-zwiepend
de ranke sloepjes, tot plots, midden in den
luchtzwijmel, de bel rinkelde, de rem-matroos
vastgreep en in smakkend gerucht de luchtvaart
gestuit stond in zachten wiegel.

—Wai blaife d’r.. blaife dr’! nog één ronde.… lekkere

metroosie, ’t is d’r soo ellendig lekker! krijschte ’n
lange meid, oranjegeel begloeid in valsch
flambouwlicht, smakkend met tongpunt paljassig d’r
heeten mond uit.

—Nog éénmaal dames, guitigde ’n mooie

matrooshelper, met glurende oogen lachend tegen de
knappe meiden, zoo maar voor ’t uitzoeken. Snel de
breed-bekraagde matroosjes, sprongen weer achter
de bootjes, duwden tegen ’t achterwerk der meiden,
dat ze gierden en raasden van opwinding om de
kriebelige duwen.—

En weer gingen in langzamen opzet, tegen elkaar, in

vaart-zwiep de ranke sloepjes, schoten ze ver buiten
tentfront de lucht in, boven het vloei-flakkerige
stangenlicht, dat als ’n toortsenrij ’t spul in z’n bont-
geel façade-geschitter bevlamde in helle-gloed. En
binnen in, langs de rood gevlagde wanden, bruiste
licht, licht, stond koppenwoeste drom meiden en
kerels, òp en neer, in storm en slinger; brasten de
sloepen door een brand-vlammigen nevel van
schijnsels. Dolle silhouetten schimden langs muren en
tentdoek, en midden in knarsend geruk, giegel en
gekrijsch, bleef dreun-deunen ’t orgel, z’n kanaljeuzen
 strot òpen, waaruit éénzelfde trompettenschetter
verbrulde.—’t Kinder-matroosje aan den ingang,
roffelde voor ’t orgel-gebouw op z’n trommel, roffelde
ròffelde uit, de razende passie der schommelmeiden
en kerels, één dof-rollende dreun van grommingen.
[292]Eindelijk klonk de bel weer, smakte sloepjes-
luchtval terug.—Dirk en Geert strompelden dronken
en waggelend uit, en al de meiden aemechtig lieten
zich de bootjes uittillen door de opgedirkte knevel-
guitige matrozen, die gniepig streelden en knepen, als
felle kereltjes lonkten en tastten.

Dansend bijéén op de estrade, roffel-hakten en

stampten de Grints, Hassels en Rink de tent-trap af.
Henk, Klaas Koome en nog ’n paar makkers stormden
áán, ieder met ’n meid, die telkens wisselden van
vrijers.

Mooie Marie Pijler, prachtslanke blonde furie,

kankaneerde vóór den stoet ’n woesten dans, met één
been de hoogte in kapriolend, haar rokken kniehoog
opgerukt, onder krampigen zwenk van ’r lijf. De
Grintjes en verblufte kerels om haar, op de tent-trap,
klapperstampten en joolden mee van pret.—Maar
Hazewind alleen sprong in sluipende lenigheid vlak
voor de furie, danste ’n vervariëerde horlepijp met
slappe kuiten en slappe polsen, als ’n Engelsche
komiek dof klapperend met z’n zolen op ’t hout. De
meid Pijler draaide langzaam, één been al hooger
uitgerekt om ’m heen, in verwilderde oogen-extase.
Haar blonde haartooi vlamde òp in gouden rossigheid,
volgedrupt van flambouwenrood, en ’r zwijmeltronie, in
wondre omstraling van harenglans, begon plots te
proesten in helsch geschater. ’t Woelde om ’t stoetje,
de luidruchtigste lachende zangers van heel de
luchtschommeltent.

De Grintjes en Hassels, nu mèt Marie Pijler

bijééngegroeid, juichten plots, toen ze ’n endje van de
estrade, in ’t helle elektriek, Guurt Hassel oppikten;
Guurt, die alleen had staan te koekeloeren naar d’r
sekretarieheertje.—Jan Grint pakte ’r vast, lolde er
opgewonden meè. Aan één rij, in slingergang, dwars
door aanstormende en weghotsende stoeten,
kankaneerden ze de Haven àf.—Plots kwam golvende
massa opdeinen uit steegjes, die meesleurde ’t
stoetje, en al de kerels en meiden weer terugdrong en
neerwierp vóór de luchtsloepen, de plek waarvan ze in
zwier en zang, afgehost waren. Dat maakte Dirk en
Rink nijdig, en woester stortten ze zich in den
kermiswoel, sleurden de meiden in armenknel en dolle
vaart mee, werkten en trampelden [293]rond, in slag en
duw, waar ze wezen wilden.—Maar de kerels hadden
geen lol genoeg op straat. ’t Was nog te stil overal. D’r
zat nog geen steigerende furie in. Hazewind schold op
dà’ terain.. van dà’ tie konsteteerde van dà’ d’r gain
duusend minse nog enterd woare.… dà’ de heule
kermisbeweging in ’t honderd liep.…—

De meiden gilden, lalden, dat ze dan maar de kroeg of

de danshuizen moesten bestormen. De neven Hassel,
warm, opgeblazen, hijgend van hitte, en verdoofd van
orgeldreun-razernij konden ’t best vinden, zeiden lievig
tot mâlkaar dat ’t toch beter zóó was dan je met
gladden smoel voorbij te loopen, zonder woord. Ze
zeiden ’t maar, wijl ze voelden dat anders Geert er van
door zou schieten met ’n ander; Geert die beefde voor
ruzie. Dàn zeilde Jan Hassel bij Pijler, dàn Piet bij Cor,
dan Henk bij Annie, in stoerigen wissel van lijven en
zoen-wangen. De blonde furie Marie duivelde en
sprong vooruit, schreeuwde in vlammige dolheid als ’n
bezetene, in hysterische stemme-krijsch kanaljeuze
liedjes uit; holde en klauterde tusschen tenten en
spullen vooròp met den dikken takstronk van alles-
zoenenden Henk, den stoet betamboereerend.
Telkens schakelde hun slingerrij stuk, door alleen-
zwalkende dronken zwervers, die aansliertten en
rondrumoerden met stuipige gebaren, kokhalzend en
verspuwend brallende liedjes. Door de hosrijen
strompelden ze heen, in beestigen slemp weer
tusschen de donkere tentruggen verwaggelend hun
kaduke lijven.—

Tegen twaalf uur, bij losbreking van schouwburg en

Café-chantants van Baanwijk en Bikkerstraat, kwam in
helle-rumoer ’n zang-stoet, galm-rauwend de haven
overhossen, dat heel ’t Hassel-Grint-groepje en
aanhangsels overgolfd verstikte in de machtige
uitbarsting van elkaar-kruisenden menschenhos,
aangolvend, al dreunender en krijsch-zwellender uit
duisteren pakhuis-havenhoek bij polderweg. Drie uur
had die uitgolvende bende opgesloten gezeten, als
muizen in ’n val, en nu plots roerde er ’n stuip van
loswoelend herleven, voortschokkend en in branding
klotsend de heele kermisjool.

Na ’n half uur waren Dirk en Willem, Henk en Rink de

meiden [294]kwijt; vonden elkaar eindelijk weer,
tusschen zwirreling van hossers en zangers. En heel
 den nacht zwierden ze van danskroeg naar
danskroeg, van herberg naar herberg, tot de Grints en
Hassels, stombezopen tegen elkaar aanvielen, lam-
gekankaneerd en lodderig zwaar, elkaar bijna niet
herkennend. De neven begonnen te krauwelen, de
meiden gierden. Marie Pijler drensde om ’n
jongenspak, sleurde zich d’r rokken van d’r lijf.
Eindelijk kwam politie die ’t dronken stelletje, achter
andere troepjes áán, van ’t kermis-terrein verduwde.

Zoo strompelden de meiden en dronken knapen, in

heesch-schreienden zang, de stik-duistere laantjes in,
om Wiereland en Duinkijk.—

[Inhoud]

III.

Ouë Gerrit had stierlijk ’t land, vloekte tegen Guurt en

zijn wijf dat de kerels ongehoord verzopen en
geradbraakt ’t huis inzwalkten. ’t Landwerk lag braak.
Tegen den ochtend waren de jongens weer op de
beenen gescharreld, stond Dirk met dikke oogen en
slappe beef-knieën tusschen de sperzieboonen,
gaaploeiend wat vruchten te zoeken in zoeten
luierrengel. Z’n oogen vol tranen gewaterd, branderig
en katterig door ’t heele lijf, loomde en sleurde ie zich
voort. Telkens op ’t pad geknield, onder ’t hoog boven
z’n hoofd dichtgegroeide ranken-groen van boonen,
zakte hij ronk-zwaar en lenden-lam in, zonder
 weerstand, met al slappere slaapgloeiing en zoete
matheid in de knuisten. Zoo, met z’n wit-blonden kop
scheef tegen de rijzen en latten van z’n boonen,
voelde ie straks te zullen neerzinken in màf, op den
zonnenden gloeigrond.

De zon vlamde doòr ’t groen; ’t zand dampte hette uit.

Eén soezerigheid was over Dirk’s kop geduizeld. De
lucht had hem eindelijk heelemaal te pakken, en eer ie
’t goed wist lag z’n heele korpus, slaap-gebogen en
verzonken, dwars tegen ’t rijzenhout. Hij ronkte en
snoffelde als ’n varken.—’n Laan achter ’m lag ’n
plukker die ook den heelen ochtend al had
tegengeworsteld, [295]maar door de snikhitte overmand
was. Het gebukt zoeken naar jonge schepseltjes had
z’n verzopen en afgetobd lijf heelemaal tol-duizelig
gemaakt en de stille invretende Augustus-brand,
schroeide nu rondom de slaapkoppen op de
blakerende droge akkers.—Ouë Gerrit nuchter en bits,
de kermisweek vervloekend bij elken stap, botste op
den ronkenden Dirk, in ’t boonengroen.—

—Moar god-liefe-Hair! hai je ’t ooit sòo sout gaite..

hoho! Dir-rik! Hai hoo! Dir-rik! schud-schreeuwde ie
woest, bonzend tegen z’n gelen kop.—

Dirk schrok, kéék òp, z’n slaperige tronie onbewust

heffend in brandende zonnesteeken, dat ie plots
bukte, blindgegooid met licht. Z’n makker, door krijsch
van den Ouë ook opgeschrikt, greep lukraak naar de
ranken, angstig-gissend of ie wèl of niet gesnapt was.
—Wat gauw kwam toch die zoete luiheid door z’n
polsen en handen gestroomd. Nou had ie ’n suizend
gevoel in zich, of ie uren òp één stuk van z’n been had
gezeten. En Dirk voelde zich branderig alsof ze’m
stukken van z’n romp hadden afgezaagd.—.…
f’rdomd.. aa’s tie nie oppaste.. gong ie wèèr.. z’n
oogen loken àl en de Ouë, stom in woede, keek maar.

—Wâ is d’r an ’t handje Ouë, lijmerde hij eindelijk

kregel uit, wa nou?

—Moar main kristis! da goan tug te waid.… Hep jai d’r

dan heuldegoar gain koorakter!.. Piet lait d’r half-dood
veur de andaifie huhu! en de moffeboone legge te
wachte.. Main Jesus.. de hellepers sakke d’r glad-
wèg! hoho! ikke mot d’r tug laifere! Enne.. ikke.. mit
main ouë bast.. lap ’t vast nie allainig! eénmoal..
andermoal.. woar mot dâ haine! dâ haine! die
moffeboone stoan d’r puur te rotte op de grond hee?—

—Nou seur d’r soo nie! kom bai de huur t’regt! wat jou
Ouë? Murge is d’r Sondag.. enne moandag doene d’r
wai vast van sellefers niks.… vàst niks! dat weut je …
is d’r kerremis-moandag.… Mô jai d’r ook moar-rais
hain trekke.… omdá niet-en-kan? Och! loat ’t ouë bier
moar werreke!

Brommend en vloekend ging Ouë Gerrit wèg,

radeloos, toch [296]stom van angst. Wel had ie fellen
lust in ’m om te gaan kijken, maar hij was d’r doodelijk
bang voor z’n eigen natuur dáár. Voor vier jaar had ’m
zoo’n tentbaas juist betrapt op ’n klein gapperijtje. Hij
had den kerel ’t vijfdubbel betaald en die had zich toen
stom gehouden. Maar uit schaamte en angst voor dien
vent, had ie in vier jaar tijd nooit meer ’n stap op de
kermis gedaan.—En ’t was zoo glad van de hand
 gegaan dat ie ’r eigenlijk nooit meer om dacht, en ’t
ook nimmer meetelde als ’n snappertje.—Want ’t
jeukte, kriebelde in ’m in de handen. Wat ie daar al
niet zien had? Hij zou zich niet weten te houên.…
Hoho … al die spulle.… die kleure.… die glommige
dingies. Sou die ’t tug-en-doen? Sou die ’t t’met
woage? Dur woa’s vast niemand die ’t wist! Most dus
puur tug ’n toeval weuse.. aàs tie die vint dur nog
antròf. Hoho.. hai waa’s dur soo malgroag.… Soo
malgroag! En ’t waa’s dur de heule eerste kair van s’n
laife daa’t ie op ’n klainighaidje betrapt wier!.… Aa’s
dur nou tùg nies van ’t land t’regt kwam, most hai s’n
aige dan hier allainig mit ’t waif doodkniese! arm,
doodarm waa’s tie tug! S’n aige brokkie grond waa’s
dur tug lang nie meer van sain. Hij poerde moar veur
’n aêr! Enne had ie dan nie s’n spulle? Wa kén d’r sain
de heule mikmak skaile? Most tie d’rof van sain grond,
wel nou! dan d’rof! hoho! F’rduufeld.. hij sou d’r nog
rais fleurig haingoan kenne.… En dan figilaire op de
klaintjes.. Hai sou d’r nou hain! Most d’r moar van
komme waa’t wou! Nou de kerels tùg de boel verloope
lieje … moar dan sou hài Sondag op stap goane …
hoho!.. Sondagòafed.. aa’s ’t drokst waa’s.. ’t dolst
t’met.. daatie nie in de lampies liep!

Brommerig en toch half voldaan dat ie ’t met zich zelf

eens was geworden klomperde hij ’t land àf, voor z’n
leegen rommel staan blijvend. Piet hurkte bij de kolen.
—

—Da koolgoedje gaif je tug moar smerige sintjes Ouë!

mos je moar nie meer sette! ikke sien d’r vast nies in!
 —Hoho.. vier-en vaif en nie genog! d’r valt t’met niks
meer te sette, stotterde Gerrit verlegen, verbaasd Piet
weer zoo frisch en flink te zien poeren, hurkend
doorzwoegen of d’r niks met [297]’m gebeurd was. De
knaap had stiekem z’n heelen kop onder de pomp
geduwd, dat ’t water langs zijn wangen droop, z’n
haren plakten en sluikten langs de slapen. Nou voelde
ie zich weer doorrild van frischheid, galmde ie z’n
kermislied uit, met schorren klank in de zonnende
ruimte rondom.—De Ouë wègklomperend, hoorde nog
lang achter z’n rug Piet’s stem:

—Oooaauw wat ’n Ska-ande,

Loage wroak van En-ge-land.

Kermis woelde door, en gloedkrateriger stond in den

avond ’t stedeke in brand, laaide ruischend z’n ros-
gloeiende flambouwenhel òpen, waarin demonisch-
donkere stoeten uit en achter tentduister en
vlammenlicht inhosten, met woest-duistere sprongen
en gebaren van kannibalen om nachtvuren.

Toch met den Zondag, kwam pas ’t èchte

kermislawaai, van alle plaatsen tegelijk. Op den
middag,—uit de treinen, uit tentwagens, uit reus-
bakkige bolderende Jan-Pleziers, lodderig als
melancholieke tufs-tufs,—drongen de stoeten áán,
van Lemper, Kerkervaart, Duinkijk, Zeekijk, Overschie,
van overal.—

Heel ’t omliggend platteland was leeggeloopen en

opgestoet naar de Wierelandsche kermis. Vee- en
kaasboeren met schom’lige vervette buiken, tuinders,
kweekers en zeelui van Dijkland en al ’t landvolk,
 dromden dooréén. Pracht-omsjaalde bevlagde
boerinnetjes, met d’r kleurgrillige mantels, bontvervige
lijfjes, rokken en vonkend gouden koppen, verdrongen
elkaar, hortend, stoeiend en woelend in zwirreling van
bloote armen, bontkleurige schouders, lachende
tronies, blank en glimmend; omlegerd van
schoremzootjes, saamgeplet met dronken, uitzinnig
krijschende zwierders, en teruggemalen tusschen
aanzwellenden kruisgang van al nieuwe kermisjolers.
—

Zoo duizenden en duizenden, op den middag al,

verwoelden in ’t enge ruim van Haven en Baanwijk.
Kleur-zwirrelende menschenzee golf-deinde en
klotste, bruiste en schuimspatte rondom, tusschen de
pronkspullen en tenten, kramen en draaimolens.—

Tegen den avond pas bulderde áán, ’t groot-geweldige

donk’re [298]rumoer, verklonk ’t stedeke in daverend
hellegerucht, gloeide en schuimflitste de kleurwoel,
donderden de bas-kelen uit hosstoeten, vergilden de
passie-meiden hun hysterischen krijsch-jubel, in
snerpenden scheurend fellen klankenmartel er
doorheen.

De lucht boven Baanwijk-breede boulevard hing laag

en starreloos-duister. In ’t diepe midden, den poortigen
allée-weg, achter de wafelkraam-ruggen, hing ros-
gouden damp, als van ’n uitbrekenden brand, angstig
moordrood in revolutie-nachten, heel den hemel daar
in gloed zengend, bang verrood van vlammen,
verborgen nog lekkend in kraterigen smeul en
windflakker. Hellegloei nevelde boven den boulevard
in de rondom duistere lucht.

Van ’t Stationsplein àf gezien, gloeide daar in

mysterisch toovervuur, de heele Baanwijk als een
verre, diepe laan, waar doorheen zwirrelden en
wemelden donk’re menschenstoeten, telkens èven en
anders weer aangegloeid in fakkelbrand en
toortsenhel, walmend gloedrood, helgeel en oranje
damp, vurig dooreenvloeiend voor tenten en façades.
En tusschen de zwart-ademende lampetten uit, van
stalletjes en kraampjes, doken òp van allen kant,
vergroot en verwilderd, donk’re woelmenschen in
sidderenden nevel; vergroeiend daarin tot titanische
wezens, met geweldszwaai van hun belichte armen,
den allegorischen gloed van hun vurige oranjekoppen,
en de geel-bronzen verflakkering van kermis-
hellesfeer die ze omzoog, omtrilde.—

Plots verdwenen de stoeten tusschen duistere mooten

en inhammen van tentruggen, ’n end dieper den
boulevard òp weer aanrossigend, onder ’t krijsch-wilde
alarm van hun kermiszang; verdreunend in kudde-
trampel, langs en om begloeide boomstammen, die
zelf verwoest stronkigden in hun schors-kervige
wildheid.

Zoo, de stoeten steigerden tusschen de verlichte

gevaarten-stammen; schuifelende troepen, wadend
door een vàl van elektrisch geflakker, blauwpaarsige
siddersfeer, dampend van boom tot boom, tent tot
tent. En daar achter weer, in duisterig diep,
overstroomd plots van avondkleuren, de woeste