Human amniotic membrane as a chondrocyte carrier vehicle/substrate: In vitro study

Abstract

Human amniotic membrane (HAM) is an established biomaterial used in many clinical applications. However, its use for tissue engineering purposes has not been fully realized. A study was therefore conducted to evaluate the feasibility of using HAM as a chondrocyte substrate/carrier. HAMs were obtained from fresh human placenta and were process to produced air dried HAM (AdHAM) and freeze dried HAM (FdHAM). Rabbit chondrocytes were isolated and expanded in vitro and seeded onto these preparations. Cell proliferation, GAG expression and GAG/cell expression were measured at days 3, 6, 9, 12, 15, 21, and 28. These were compared to chondrocytes seeded onto plastic surfaces. Histological analysis and scanning electron microscopy was performed to observe cell attachment. There was significantly higher cell proliferation rates observed between AdHAM (13-51%, P=0.001) or FdHAM (18-48%, p = 0.001) to chondrocytes in monolayer. Similarly, GAG and GAG/cell expressed in AdHAM (33-82%, p = 0.001; 22-60%, p = 0.001) or FdHAM (41-81%, p = 0.001: 28-60%, p = 0.001) were significantly higher than monolayer cultures. However, no significant differences were observed in the proliferation rates (p = 0.576), GAG expression (p = 0.476) and GAG/cell expression (p = 0.135) between AdHAM and FdHAM. The histology and scanning electron microscopy assessments demonstrates good chondrocyte attachments on both HAMs. In conclusion, both AdHAM and FdHAM provide superior chondrocyte proliferation, GAG expression, and attachment than monolayer cultures making it a potential substrate/carrier for cell based cartilage therapy and transplantation.

Direct comparison of the potency of human mesenchymal stem cells derived from amnion tissue, bone marrow and adipose tissue at inducing dermal fibroblast responses to cutaneous wounds

“…In a mouse model of excisional wound healing, we injected equal numbers of labeled hAMSCs, hBMSCs or hADSCs around the wound to compare the therapeutic effects of the three types of cells. HAM has been used as a dressing for surface wounds based on its anti-inflammatory and antimicrobial effects (35)(36)(37)(38)(39)(40). Our studies revealed that hADSCs significantly reduced the wound size and accelerated re-epithelialization from the wound edge, suggesting that hADSC transplantation is a more potent therapeutic treatment for skin wounds than hBMSC or hAMSC transplantation.…”

Concise Review: Fetal Membranes in Regenerative Medicine: New Tricks from an Old Dog?

“…In their lyophilized state, decellularized AM showed potential as a novel barrier for guided bone regeneration in large bone defect where the membrane could afford protected space for osteogenesis while concurrently protecting against fibroblast invasion . Furthermore, an extract of AM has been postulated as an anti‐hemolytic and anti‐thrombotic due to its abundance in glycosaminoglycans (i.e., perlecan and hyaluronic acid) which inhibit coagulation, as well as as IL‐10 and MMP‐9 which inhibit platelet aggregation .…”

“…The fetal membranes have been used in other applications such as their utility as a feeder layer for human embryonic stem cell derivation and maintenance [103]. Air-dried and freeze-dried amniotic membranes (AM) have been assessed for their feasibility as a growth substrate for chondrocyte expansion, and were found to improve chondrocyte proliferation, GAG expression and attachment than monolayer cultures [104]. In their lyophilized state, decellularized AM showed potential as a novel barrier for guided bone regeneration in large bone defect where the membrane could afford protected space for osteogenesis while concurrently protecting against fibroblast invasion [105].…”

Amniotic membrane as a scaffold in wound healing and diabetic foot ulcer: an experimental technique and recommendations

“…9,10 Furthermore, amniotic membranes have also shown promising results as nerve wrap, in cartilage restoration, osteoarthritis and in tendon repair. [11][12][13] So far, various methods have been explored to preserve amniotic membrane such as fresh storage (store at 4°C), cryopreservation (either in minus 86°C or liquid nitrogen), freeze drying, and dehydration, each having the objective of to retain all components of the tissue as close as to fresh tissue and have longer shelf life. Each method is having its own pros and cons.…”