Abstracts / Cytotherapy 23 (2021) 1 39
availability when compared with other sources. However, several
factors may compromise ADSCs‘ genetic integrity, such as the ana-
tomical location of the adipose tissue (AT), cell culture expansion,
and environmental stressors such as UVB radiation. Loss of genetic
integrity is associated with a decrease in cell quality for experiments
and therapeutic applications. Therefore, in this study, we compara-
tively evaluated the presence of g -H2AX, a marker of DNA double-
strand breaks, in human facial and abdominal ADSCs during long
periods of cultivation under standard conditions or exposed to UVB
radiation.
Methods: ADSCs were obtained from female donors of facial AT (N=3)
and abdominal AT (N=3). We evaluated the presence of g -H2AX in
facial and abdominal ADSCs in early (P2-P5), medium (P12-P18), and
late (P25-P35) passages, under standard culture conditions and after
48 hours of exposure to 30 mJ/cm2 UVB.
Results: g -H2AX expression gradually increased in both cell types
under standard culture conditions. Facial and abdominal ADSCs simi-
larly expressed g -H2AX in early (15.1 § 14.6% and 14.8 § 10.4%,
respectively) and late passages (61.1 § 20% and 73 § 13.5%, respec-
tively). In medium passages, the presence of g -H2AX was significantly
greater in facial than in abdominal ADSCs (P = 0.019). Upon UVB radia-
tion, g -H2AX was significantly 1.3 times and 1.2 times higher in
abdominal ADSCs in the early and medium passages, respectively
(P = 0.01), but both cell types showed similar late passages levels (90%
and 96% in ADSCs facial and abdominal, respectively).
Conclusion: Our findings indicate that DNA damage occurs similarly
between facial and abdominal ADSCs under standard culture condi-
tions. However, UVB radiation generates a greater increase in DNA
double-strand breaks in abdominal rather than facial ADSCs, suggest-
ing that facial cells might be more resistant to UVB radiation. Future
studies will further investigate if facial and abdominal cells respond
differently to environmental stressors.
45
HISTOLOGICAL AND MICROBIOLOGICAL EVALUATION IN MICE OF
CYANOACRYLATE (HISTOACRYL) AS POSSIBLE REGENERATIVE
AGENT TO REPLACE TRADITIONAL SUTURES
n3, P. Barba4, A. Chiliquinga1,
A. Villagomez1,2,*, T. Borja3, P. Ponto
I. Yambarela4, C. Pupiales4, D. Suquillo5,6,7, R.F. Díaz1,6, G. Segnini1,2,
A. Caicedo6,7,8,9, F. Cabrera1,6,7
1
Escuela de Medicina Veterinaria, Universidad San Francisco de Quito
USFQ, Quito, Ecuador, 2Servicio de Cirugía de Alta Complejidad y Mín-
ima invasio n - Hospital Docente de WOUND HEALING AFTER ONCO-
LOGICAL SURGERY, Especialidades Veterinarias, Universidad San
Francisco de Quito USFQ, Quito, Ecuador, 3Servicio de Patología, Hos-
pital Voz Andes, Quito, Ecuador, 4Carrera de Biotecnología, Universi-
dad Te cnica del Norte, Ibarra, Ecuador, 5Ingeniería en Procesos
Biotecnolo  gicos, Colegio de Ciencias Biolo  gicas y Ambientales
COCIBA, Universidad San Francisco de Quito USFQ, Quito, Ecuador,
6
Instituto de Investigaciones en Biomedicina, Universidad San Fran-
cisco de Quito USFQ, Quito, Ecuador, 7Mito-Act Research Consortium,
Quito, Ecuador, 8Escuela de Medicina, Universidad San Francisco de
Quito USFQ, Quito, Ecuador, 9Sistemas Me dicos SIME, Universidad
San Francisco de Quito USFQ, Quito, Ecuador
Background: Traditional sutures, staples and surgical tapes are the
most common methods used to close wounds in veterinary patients.
However, these methods can induce granuloma and fistulas forma-
tion, loose sutures and wound leaks. Lately, tissue adhesives such as
cyanoacrylates (HistoacrylÒ ) have become popular, this glue can
bond tissues such as endothelium, mucosa, skin, blood and bone rap-
idly. Studies using cyanoacrylates have shown bond strength, bacteri-
ostatic and hemostatic properties. The objective of this work is to
compare the use of n-butyl-cyanoacrylate with non-absorbable
sutures in tissue regeneration by histology and microbiological
17
analysis in mice. (All procedures were approved by the USFQ Ethics
Committee on Animal Experimentation, Doc# 2020-001).
Methods: Two cuts of 1 cm were perfomed in the dorsal skin of each
mouse: one cut was bonded by simple suture (control), and the other
with tissue adhesive (Histoacryl). Albino mice were anesthetized
before the surgical procedure; analgesics were provided in the water
after the surgery to minimize pain. After 48 hours of the surgical pro-
cedure, the mice were euthanized and biopsies of 2 cm2 were col-
lected for each cut. Three mice were used for histological evaluation
and two for microbiology tests. Assays were repeated 4 times. For
microbiology tests, samples were resuspended in 1 ml of TBS culture
broth by vortex. Colony forming units (CFU) were evaluated for two
dilutions after incubation on Nutrient Agar for 48 hours. For histolog-
ical analysis, biopsies were fixed with formaldehyde, cut by a micro-
tome (Leyca RM2155) and stained with trichrome, haematoxylin and
eosin. Statistical analysis was performed by Mann-Whitney and Krus-
kal-Wallis tests using GraphPad Prism software.
Results: No differences were found in the microbiological and histo-
logical analysis.
Conclusions: The results showed that there is no significant difer-
ence between n-butyl-cyanoacrylate and non-absorbable sutures, no
increased risk of microbiological contamination or regenerative
improvements were shown.
46
HISTOLOGICAL EVALUATION OF CHONDRAL LESION IN RABBIT’S
KNEE JOINTS TREATED WITH MESENCHYMAL STEM CELL
ASSOCIATED WITH COLLAGEN MEMBRANE
ML Barbosa1,*, GV dos Santos2, L Fracaro1, JA Villanova JR2,
S Nagashima3, APC Martins3, CBV De Paula3, SO Ioshii3,
LGA Capriglione1, PH Utumi1, LM Boldrini-Leite1, PRS Brofman1,
CLK Rebelatto1
1
Core for Cell Technology, School of Medicine, Pontifícia Universidade
lica do Parana
Cato  (PUCPR), Curitiba, Brazil, 2Graduate Program in
Animal Science, PUCPR, Curitiba, Brazil, 3Experimental Pathology Lab-
oratory, School of Medicine, PUCPR, Curitiba, Brazil
Background: The repair of chondral defects is challenging because
cartilage is avascular and has a limited potential of regeneration due
to the low mitotic activity of chondrocytes. Mesenchymal stem cells
(MSC) are immunomodulatory cells and have the potential to differ-
entiate to chondrocytes. Association of MSC and a biomaterial would
be an alternative for chondral lesion treatment. This study evaluated
the rabbit cartilage tissue, with a chondral lesion, after umbilical cord
and adipose tissue MSC transplantation, associated with a collagen
membrane.
Methods: This study was approved by the local Ethics Committee
(CAEE: 56776716.1.1001.0020). Histological and immunohistochemi-
cal analysis were performed on rabbit’s knee joints that have been
submitted to injury by abrasion and microfractures in the subchon-
dral bone. For all animals of each group (n=11/group), the collagen
membrane was applied (Group A), associated with umbilical cord
MSC (Group B) and adipose tissue MSC (Group C). Quantification of
collagen types I and III was performed using picrosirius red staining.
Analysis of type II collagen was performed by immunohistochemistry
with anti-type II collagen antibody.
Results: A higher density of collagen type I in the three groups was
observed, in contrast to collagen type III. There was no significant dif-
ference between the groups, regarding the amount of collagen (group
A x group B, p=0,4278; group B x group C, p=0,3939; group A x group
C, p=0,5622). Group B showed a higher expression of collagen type II,
although no statistical difference was observed when comparing the
three groups (p=0,111).
Conclusion: The expression of type I, III, and mainly type II collagen,
which is present in hyaline cartilage, demonstrates that the three