The superiority of the autografts inactivated by high hydrostatic pressure to decellularized allografts in a porcine model

Abstract

We are developing a novel skin regeneration therapy in which the inactivation of nevus tissue via high hydrostatic pressure (HHP) is used in the reconstruction of the dermis in combination with a cultured epidermal autograft. In this study, we used a porcine skin graft model to explore whether autologous skin including cellular debris inactivated by HHP or allogeneic skin decellularized by HHP is better for dermal reconstruction. Grafts (n = 6) were prepared for five groups each: autologous skin without pressurization group (control group), autologous skin inactivated by 200 MPa group, autologous skin inactivated by 1000 MPa group, allogeneic skin decellularized by 200 MPa group, and allogeneic skin decellularized by 1000 MPa group. All of the grafts at 1, 4, and 12 weeks showed complete engraftment macroscopically. The mean areas of the grafts of the control group (p < 0.01) and autologous 200 MPa group (p < 0.01) were larger than that of the allogeneic 1000 MPa group at four weeks after implantation. The thickness of the control group and autologous 200 MPa group was comparable, and that of the autologous 200 MPa group was significantly thicker than that of the allogeneic 200 MPa group (p < 0.01). This suggests that the autologous dermis was superior to the allogeneic decellularized dermis as a skin graft, and that HHP at 200 MPa provided a better outcome than HHP at 1000 MPa. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 105B: 2653-2661, 2017.

Exploration of the Pressurization Condition for Killing Human Skin Cells and Skin Tumor Cells by High Hydrostatic Pressure

“…High hydrostatic pressure (HHP) is a safe method of physically inactivating cells or tissues promptly without using chemicals, such as detergents, that is commonly used to prepare decellularized tissues. We previously reported that HHP at 200 MPa for 10 min was able to inactivate all cells in porcine skin, human skin, and human nevus tissue (giant congenital melanocytic nevus (GCMN), representative of human congenital skin tumor) [1][2][3][4][5][6]. Furthermore, HHP at 200 MPa did not damage the extracellular matrix, although HHP at 1000 MPa damaged and altered the epidermal basement membrane to some degree and prevented the survival of human cultured epidermis (hCE) on the pressurized skin or nevus [1].…”

“…Furthermore, HHP at 200 MPa did not damage the extracellular matrix, although HHP at 1000 MPa damaged and altered the epidermal basement membrane to some degree and prevented the survival of human cultured epidermis (hCE) on the pressurized skin or nevus [1]. In addition, we reported that autologous dermis pressurized at 200 MPa without removing cellular debris showed less contracture after grafting than decellularized allogeneic dermis using a porcine model [2].…”

The sustained release of basic fibroblast growth factor accelerates angiogenesis and the survival of the inactivated dermis by high hydrostatic pressure

“…We previously reported that HHP exceeding 200 MPa completely inactivated cells in the nevus tissue without damaging the structure of the ECM, while HHP exceeding 500 MPa damaged the epidermal basement membrane [25]. Although we found that the remaining cellular debris did not adversely affect the take of the inactivated skin [23, 24, 27, 35], the infiltration of fibroblasts from the wound bed into the inactivated skin took more than a week in a porcine autograft model, and the graft tended to be absorbed after four weeks [27, 35].…”

Repair of cartilage defects with devitalized osteochondral tissue: A pilot animal study

“…Because of its strong chondroinductive potential, we hypothesized that HHP‐treated osteochondral plugs could be used as scaffold for the reconstruction of (osteo‐) chondral lesions in vivo . Although HHP‐treated porcine skin was implanted successfully in vivo , implantation of osteochondral tissue treated with HHP is not characterized in animal models so far …”

“…Although HHP-treated porcine skin was implanted successfully in vivo, implantation of osteochondral tissue treated with HHP is not characterized in animal models so far. 18 Therefore, in this pilot study, we evaluated the feasibility of HHP-devitalized plugs to repair osteochondral defects in a rabbit model. We assumed that the natural origin of the devitalized plugs may support their integration into the adjacent tissue and represent an environment which attracts cell migration.…”