Preparation and characterization of injectable fibrillar type I collagen and evaluation for pseudoaneurysm treatment in a pig model

Choi

et al. 2012

Biotechnology Progress

Collagen, the most abundant protein in vertebrates, is a useful biomaterial in pharmaceutical and medical industries. So far, most collagen has been extracted from animals and cadavers. Herein, we suggest human adipose tissue, which is routinely abandoned after liposuction, as a plentiful source of human collagen. In this study, human collagen was obtained from adipose tissue through two successive major steps: (i) extraction of the extracellular matrix (ECM) by pulverization, centrifugation, alkaline, and alcohol treatment; (ii) isolation of collagen from ECM by pepsin treatment in dilute acetic acid. The purified human adipose-derived collagen was characterized by Fourier transform infrared spectroscopy, polyacrylamide gel electrophoresis, amino acid analysis, and circular dichroism spectroscopy. The extracted collagen showed a typical triple helix structure, good thermal stability due to abundant imino acids, and high solubility at acidic pH. The collagen greatly facilitated the adhesion and proliferation of human adipose-derived stem cells and normal human dermal fibroblasts on polystyrene plates. These results suggest that human adipose tissue obtained by liposuction can provide human collagen for use in cosmetics, pharmaceutics, and medicine.

Section: Resultsmentioning

confidence: 99%

Human collagen isolated from adipose tissue

Choi

et al. 2012

Biotechnology Progress

ACS Biomater. Sci. Eng.

“…Collagen films were sterilized as follows: GP of 58% hydrogen peroxide at 6 mg/L for 45 min diffusion and 20 min radiofrequency excitation; , GI at 8.8 kGy at room temperature; , EO at 750 mg/mL at 40 °C for 12 h, , and 70% ET for 30 min at room temperature. , Nonsterile (NS) films were used as control. The disinfection/sterilization cycle of GP and GI was ensured with a dosimeter indicator.…”

Section: Methodsmentioning

confidence: 99%

“…Gamma irradiation, for example, has been shown to cause structural changes and induce significant reduction in mechanical properties, resistance to degradation, and cell attachment. − E-beam irradiation at low and high doses significantly decreased the ultimate strain and toughness of human cortical bone (almost 80% of the organic matter in bones is collagen , ), while high doses of e-beam irradiation negatively impacted on bone regeneration of octacalcium phosphate collagen composites . Ethylene oxide has been shown to reduce stability and cell attachment of collagen-based devices. , In recent years, disinfection/sterilization through hydrogen peroxide gas plasma has been advocated, as it can be conducted at temperatures (<50 °C) well-tolerated even by non-cross-linked collagen devices and has demonstrated minimal detrimental effects in comparison to ethylene oxide disinfection/sterilization. , Ethanol/alcohol treatments are also gaining pace, as they do not affect structural, degradation, and mechanical properties. − It is, however, worth noting that alterations in pore size due to shrinkage have been reported, , and it is not used extensively in clinical settings as it cannot eliminate spores and viruses …”

Section: Introductionmentioning

confidence: 99%

“…15,20 In recent years, disinfection/ sterilization through hydrogen peroxide gas plasma has been advocated, as it can be conducted at temperatures (<50 °C) well-tolerated even by non-cross-linked collagen devices 11 and has demonstrated minimal detrimental effects in comparison to ethylene oxide disinfection/sterilization. 21,22 Ethanol/alcohol treatments are also gaining pace, as they do not affect structural, degradation, and mechanical properties. 23−25 It is, however, worth noting that alterations in pore size due to shrinkage have been reported, 24,26 and it is not used extensively in clinical settings as it cannot eliminate spores and viruses.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Influence of Cross-Linking Method and Disinfection/Sterilization Treatment on the Structural, Biophysical, Biochemical, and Biological Properties of Collagen-Based Devices

Delgado

Fuller

Zeugolis

2018

Disinfection/sterilization is an essential step during the manufacturing process of any implantable medical device. Cross-linking is also required for biopolymers to control resistance to degradation and enhance mechanical integrity. To date, there is still no single disinfection/sterilization treatment and cross-linking method that can be used universally for collagen-based devices. Herein, we assessed the influence of ethylene oxide, ethanol, gamma irradiation, and gas plasma disinfection/sterilization on the structural, biophysical, biochemical, and biological properties of self-assembled collagen films cross-linked with 4-arms polyethylene glycol succinimidyl glutarate and genipin. Microscopy analysis revealed that gas plasma treatment induced the most profound differences in the non-cross-linked and 4-arms polyethylene glycol succinimidyl glutarate cross-linked collagen films. Gas plasma also significantly increased the swelling ratio of the non-cross-linked and the 4-arms polyethylene glycol succinimidyl glutarate cross-linked films. Non-cross-linked and gas plasma treated 4-arms polyethylene glycol succinimidyl glutarate collagen films exhibited the lowest resistance to collagenase degradation and denaturation temperature. Between the non-cross-linked groups, the gas plasma treatment resulted in the collagen films with the lowest stress at break, strain at break, force at break, and Young’s modulus values. Within the 4-arms polyethylene glycol succinimidyl glutarate groups, the ethylene oxide treatment resulted in collagen films with the lowest stress at break, strain at break, force at break, and Young’s modulus values. Within the genipin groups, the gas plasma treatment resulted in collagen films with the lowest stress at break, strain at break, force at break, and Young’s modulus values. Proliferation, metabolic activity, and viability of human skin fibroblasts were not affected as a function of cross-linking method and disinfection/sterilization treatment. However, proliferation, metabolic activity, and viability of THP1 cells were significantly reduced as a function of the cross-linking method, but they were not affected as a function of the disinfection/sterilization treatment. Overall, our data illustrate that the cross-linking method and the disinfection/sterilization treatment differentially affect the structural, biophysical, biochemical, and biological properties of collagen-based devices, and thus, they should be optimized according to the clinical indication.

“…It comprised fragmented connective tissue matrix and vascular structures composed of endothelial cells and pericytes and other regenerative cells, such as the ASCs. ECM components directly influence cell behaviour, including adhesion, spreading, proliferation and migration (Geutjes et al, 2010;Kim et al, 2012;Prockop, 2007;Wang et al, 2013a), suggesting a positive effect on the cells embedded in the microtissue-SVF. Qiu et al (2018) employ decellularised fat tissue ECM as a scaffold for reseeding MSCs, resulting in improved regenerative efficacy in muscle tissue injury.…”

Section: Fig 4 Flow Cytometry Analysis Of Svf Subpopulationsmentioning

confidence: 99%

Adipose-tissue-derived therapeutic cells in their natural environment as an autologous cell therapy strategy: the microtissue-stromal vascular fraction

Nürnberger¹,

Lindner²,

Maier³

et al. 2019

eCM

The prerequisite for a successful clinical use of autologous adipose-tissue-derived cells is the highest possible regenerative potential of the applied cell population, the stromal vascular fraction (SVF). Current isolation methods depend on high enzyme concentration, lysis buffer, long incubation steps and mechanical stress, resulting in single cell dissociation. The aim of the study was to limit cell manipulation and obtain a derivative comprising therapeutic cells (microtissue-SVF) without dissociation from their natural extracellular matrix, by employing a gentle good manufacturing practice (GMP)-grade isolation. The microtissue-SVF yielded larger numbers of viable cells as compared to the improved standard-SVF, both with low enzyme concentration and minimal dead cell content. It comprised stromal tissue compounds (collagen, glycosaminoglycans, fibroblasts), capillaries and vessel structures (CD31 + , smooth muscle actin + ). A broad range of cell types was identified by surface-marker characterisation, including mesenchymal, haematopoietic, pericytic, blood and lymphatic vascular and epithelial cells. Subpopulations such as supra-adventitial adipose-derived stromal/stem cells and endothelial progenitor cells were significantly more abundant in the microtissue-SVF, corroborated by significantly higher potency for angiogenic tube-like structure formation in vitro. The microtissue-SVF showed the characteristic phenotype and tri-lineage mesenchymal differentiation potential in vitro and an immunomodulatory and pro-angiogenic secretome. In vivo implantation of the microtissue-SVF combined with fat demonstrated successful graft integration in nude mice. The present study demonstrated a fast and gentle isolation by minor manipulation of liposuction material, achieving a therapeutically relevant cell population with high vascularisation potential and immunomodulatory properties still embedded in a fraction of its original matrix.