Intestinal tissue sealing and

Intestinal adhesion prevention

Vanshika Singh
Dr. Mohammad Raisul Abedin
Dr. Mallikarjun Gosangi
Rege Bioengineering lab
05/26/2022

1
 INTRODUCTION

Intestinal tissue sealing

 ~30% patients suffer from anastomotic
leakages following intestinal surgeries.
 Can result in chronic pain, bowel obstruction,
and morbidity.
 Several approaches like secondary
enforcement of the surgical site have been
tested.
 Chitosan based material developed in the lab
for reinforcing the surgical site after suturing
is working well in rats.

Kishan, Alysha, et al., 2020

Huang, Huang-Chiao, et al., 2013 2
Chitosan film preparation
• Glycerol replaced by PVA as plasticizer
• Materials being tested for adhesivity:

1. CS-HMW
2. CS-MMW
3. CS-LMW
4. CS-Q1
5. CS-Q2
6. CS-Q3
7. CS-Cys
8. CS-GSH
9. CS-Cat
10. CS-Methacrylate
11. CS-HMW
12. Surgilux-based

3
Chitosan film characterization

• Tensile strength
• Young’s modulus
• Swelling profile
• Photothermal response
• UTS with porcine intestine
• Dynamic Mechanical Analysis (DMA)
• Ex vivo burst pressure

4
Photothermal response

Commercial chitosan Quaternized chitosan

5
Ultimate Tensile Strength

• Statistically non-significant difference between different

films compared to surgilux-based films.
• Films chosen for end-to end studies: HMW-glycerol, Q2,
Q3, and CS-MAA
6
End-to-end anastomosis
0.12

0.1

0.08
Burst Pressure (MPa)

0.06

0.04

0.02
• No. of sutures used in surgeries: 1 suture
0
ct re re e e e every 3-5 mm
t a tu tu t ur t ur t ur
In su su su su su • Circumference of intestine: 6 cm
16 14 1 2 1 0 8
• No. of sutures used: 16 sutures
• For ~50% leakage model: 8 sutures
7
End-to-end anastomosis

0.12 0.12
0.1 0.1
Burst Pressure

Burst Pressure
0.08 0.08
(MPa)

0.06 0.06

(MPa)
0.04 0.04
0.02 0.02
0
Intact 8 suture+ 0
Intact 8 suture+ 8 suture+
HMW_LASE HMW_LASE MA_LASE

CS-MA film more flexible, causing lesser stricture

8
 INTRODUCTION

Intestinal adhesion prevention

 ~90% patients develop intra-abdominal adhesions
following intestinal surgeries.
 Can result in chronic pain, bowel obstruction, and
morbidity.
 Several adhesion barriers have been developed
and commercialized eg. Seprafilm & Interceed.
 Limitations like reduced efficacy in the presence of
blood (Interceed) or delayed inflammatory-
mediated process (Seprafilm)

Kishan, Alysha, et al., 2020

https://www.fairview.org/Patient-Education/Articles/English/u/n/d/e/r/Understanding_Lysis_of_Adhesions_90784 9
 Serous membrane (or serosa)

 A smooth membrane consisting of a thin connective

tissue layer and a thin layer of cells.
 Secretes serous fluid to allow lubricated sliding
movements between opposing surfaces.
 Due to its inherent lubricating properties, might act
as an anti-adhesive agent for post-operative
adhesions prevention.
 The biomaterial derived from serosa via
decellularization process will be tested for anti-
adhesion applications.

Siri, Saeed, et al., 2020

10
Serosa. (2020, August 13). https://med.libretexts.org/@go/page/8040
Preparation of Decellularized Serosal Hydrogel
(‘Serogel’)

11
Mechanical characterization

 Adhesion property of Serograft on porcine skin and small intestine ex vivo (90-

degree peel test)

 Rheological testing (Serogel viscoelastic modulus, Gelation kinetics of Serogel and

turbidimetric kinetics)

 UTS/ DMA analysis of Serograft (undigested), Seprafilm (positive control)

12
Tensile strength (serograft)

20
18
16
14

Max Force (N)

12
10
8
6
4
2
0
A#4 (1 layer) A#4 (2 layer) A#5 (1 layer) A#5 (2 layer)

13
 Adhesion testing (serograft)

0.6

Work of Adhesion (N.mm)

0.5

0.4

0.3

0.2

0.1

0
Chitosan film Seprafilm A#4 serograft A#5 serograft
(HMW)

Seprafilm because of water absorption forms hydrogel & sticks to the tissue
Smaller load cell needed due to very low force values & noise in data
https://www.researchgate.net/publication/330664104_Mechanical_characterization_and_ex_vivo_evaluation_of_anticancer_and_antiviral_drug_printed_bioadhesive_film_for_the_treatment_of_cervical_cancer 14
Rheological testing

DHR-20, TA instruments (digested serosa-serogel)

Material tested: (Animal #2)
 4 mg/ml Collagen (control)
 4 mg/ml Serosal Hydrogel
 10 mg/ml Serosal hydrogel
 15 mg/ml Serosal Hydrogel
Temperature Settings: Temperature ramp
from 22-40°C for approx. 10 min  

15
Rheological testing

DHR-30, TA instruments (digested serosa-serogel)

 Material tested: (Animal #3)
• 15 mg/ml Serosal Hydrogel (digested)
 Temperature Settings: Temperature ramp
from 22-37°C for 10 min  

16
Rheological testing

DHR-30, TA instruments (undigested serosa-serogel)

Material tested: (Animal #3)
15 mg/ml Serosal Hydrogel (undigested)
Temperature Settings: Temperature ramp
from 22-37°C for 10 min  

17
Rheological testing

Anton Paar rheometer: Animal #2 (digested serosa-serogel)

Response of 15 mg/ml hydrogel (G' and G")
to an oscillatory temperature ramp from
22-37°C , constant temperature at 37°C and
again and ramp from 37-50°C

18
Gelation Kinetics
Turbidimetric assay
Material tested: (Animal #1, n=1)
 4 mg/ml Collagen
 4 mg/ml Serosal Hydrogel
 6 mg/ml Serosal hydrogel
 8 mg/ml Serosal Hydrogel
 10 mg/ml Serosal Hydrogel
 15 mg/ml Serosal Hydrogel
Temperature Settings: 37°C

19
Gelation Kinetics
Turbidimetric assay Material tested: (Animal #2, n=1)
 4 mg/ml Collagen
 4 mg/ml Serosal Hydrogel
 6 mg/ml Serosal hydrogel
 8 mg/ml Serosal Hydrogel
 10 mg/ml Serosal Hydrogel
 15 mg/ml Serosal Hydrogel
Temperature Settings: 37°C
Material tested: (Animal #2, n=2)
 4 mg/ml Collagen
 4 mg/ml Serosal Hydrogel
 6 mg/ml Serosal hydrogel
 8 mg/ml Serosal Hydrogel
 10 mg/ml Serosal Hydrogel
 15 mg/ml Serosal Hydrogel
Temperature Settings: 37°C
20
Gelation Kinetics
Visual observation

21
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