Preparation, characterization, and xenotransplantation of the caprine acellular dermal matrix

Abstract

Background Caprine skin is a promising biomaterial for tissue‐engineering applications. However, tissue processing is required before its xenogenic use. Aims Therefore, the purpose of this study was to evaluate the structural integrity and biocompatibility of the caprine skin after de‐epithelialization, using sodium chloride (NaCl) and trypsin solutions, followed by de‐cellularization using sodium dodecyl sulfate (SDS) solution. Materials & Methods The caprine skin was de‐epithelialized using NaCl (2‐4 mol/L) and trypsin (0.25%‐0.5%) followed by the treatment of SDS (1%‐4%) solution over a period of time. Acellularity of the prepared matrix was confirmed histologically and characterized by appropriate staining, scanning electron microscopy (SEM), DNA quantification, and Fourier‐transform infrared (FTIR) spectroscopy. The caprine acellular dermal matrix (CADM) was used for the repair of spontaneously occurring abdominal hernia in ten buffaloes. The biocompatibility of the CADM was evaluated using clinical, hematological, biochemical, and anti‐oxidant parameters. Results Histologically, the skin treated with 0.25% trypsin in 4 mol/L NaCl for 8 hours resulted in complete de‐epithelialization. Further treatment with 2% SDS for 48 hours demonstrated complete acellularity and orderly arranged collagen fibers. The SEM confirmed a preservation of collagen arrangement within CADM. The DNA content was significantly (P < .05) lower in CADM (46.20 ± 7.94 ng/mg) as compared to fresh skin (662.56 ± 156.11 ng/mg) indicating effective acellularity. The FTIR spectra showed characteristic collagen peaks of amide A, amide B, amide I, amide II, and amide III in CADM. All the 10 animals recovered uneventfully and remained sound. Hematological, biochemical, and anti‐oxidants findings were unremarkable. Conclusion Results indicated the acceptance and biocompatibility of the xenogenic caprine acellular dermal matrix for abdominal hernia repair in buffaloes without complications.

Bubaline Diaphragm Matrix: Development and Clinical Assessment into Cattle Abdominal Hernia Repair

“…In the present study, a non-significant (P > 0.05) change was observed in GSH activity and catalase levels of animals, indicating absence of oxidative stress following implantation. Similar findings were also reported in buffaloes after xenogenic application of caprine acellular dermal matrix [28].…”

Xenotransplantation literature update, July/August 2020

“…Fibrin‐induced angiogenesis was observed when transplanted subcutaneous, which enhanced NPI graft survival and long‐term function in the subcutaneous space. The other manuscript from Kuppan et al focused on human adipose‐derived mesenchymal stem cells (AdMSCs) 17 . A co‐transplantation of human AdMSCs (1.0 × 10 6 ) with NPIs (3.0 × 10 3 ) within the prevascularized subcutaneous space achieved earlier normoglycemia, improved glucose tolerance, superior stimulated serum porcine insulin, and increased graft insulin content compared to NPI transplantation alone in immunodeficient B6.129S7‐Rag1tm1Mom/J mice.…”

Fourier Transform Infrared Spectroscopy of the Animal Tissues

“…The extracellular matrix (ECM) scaffolds primarily composed of structural collagen protein are widely used in tissue engineering and regenerative medicine [1][2][3][4][5][6][7][8][9][10][11][12][13][14][15]. These are usually prepared from animal tissues by decellularization process.…”

“…Enzymatic techniques of decellularization include the use of protease (trypsin) [1-5, 8, 10, 12-15], endonucleases and exonucleases. Chemical methods of decellularization include the use of acids and alkalis (acetic acid, peracetic acid, hydrochloric acid, sulfuric acid, ammonium hydroxide), nonionic detergents (Triton X-100), ionic detergents (sodium dodecyl sulfate, sodium deoxycholate, Triton X-200) [1][2][3][4][5][6][7][8][9][10][11][12][13][14][15], zwitterionic detergents (3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate, sulfobetaine-10, sulfobetaine-16), organic solvent (Tri(n-butyl)phosphate) [3,10], hypertonic and hypotonic solutions [2,3,8,10,13,15], and chelating agents (EDTA). These reagents at higher concentrations extensively disrupt the structural proteins of ECM scaffolds and make it impossible to analyze by routine techniques [17].…”

“…These reagents at higher concentrations extensively disrupt the structural proteins of ECM scaffolds and make it impossible to analyze by routine techniques [17]. Fourier transform infrared (FTIR) spectroscopy is one of the preferred technique for identification of biomolecules through the study of their characteristic vibrational movements [11,13,14,18]. This technique is simple, reproducible, nondestructive to the tissue, and only small amounts of tissue (micrograms to nanograms) with a minimum preparation are required.…”