Effect of Decellularized Extracellular Matrix Bioscaffolds Derived from Fibroblasts on Skin Wound Healing and Remodeling.

The mammalian tissue extracellular matrix (ECM) has been used as a scaffold to facilitate the repair and reconstruction of numerous tissues. However, the material properties of decellularized ECM (dECM) from in vitro cell cultures and the effect of these properties on wound remodeling remain unclear. To elucidate its biological activity, we extracted dECM from human lung fibroblasts, fabricated it into a patch, and applied it to a full-thickness skin wound. The fibroblast-derived dECM (fdECM) maintained the content of collagen Ⅰ, collagen Ⅳ, and elastin, and the extraction process did not damage its critical growth factors. The fdECM-conjugated collagen patch (COL-fdECM) facilitated wound contraction and angiogenesis in the proliferative phase when applied to the in vivo full-thickness skin wound model. Moreover, the COL-fdECM treated wound showed increased regeneration of the epidermal barrier function, mature collagen, hair follicle, and subepidermal nerve plexus, suggesting qualitative skin remodeling. This therapeutic efficacy was similarly observed when applied to the diabetic ulcer model. fdECM was shown to help remodel the tissue by regulating fibroblast growth factors, matrix metalloproteinases, and tissue inhibitors of metalloproteinases via the p38 and ERK signaling pathways in an in vitro experiment for understanding the underlying mechanism. These results provide a biological basis for cell-derived ECM as a multi-functional biomaterial applicable to various diseases.

In Vitro Characterization of a Novel Human Acellular Dermal Matrix (BellaCell HD) for Breast Reconstruction.

In the past, acellular dermal matrices (ADMs) have been used in implant-based breast reconstruction. Various factors affect the clinical performance of ADMs since there is a lack of systematic characterization of ADM tissues. This study used BellaCell HD and compared it to two commercially available ADMs-AlloDerm Ready to Use (RTU) and DermACELL-under in vitro settings. Every ADM was characterized to examine compatibility through cell cytotoxicity, proliferation, and physical features like tensile strength, stiffness, and the suture tensile strength. The BellaCell HD displayed complete decellularization in comparison with the other two ADMs. Several fibroblasts grew in the BellaCell HD with no cytotoxicity. The proliferation level of fibroblasts in the BellaCell HD was higher, compared to the AlloDerm RTU and DermACELL, after 7 and 14 days. The BellaCell HD had a load value of 444.94 N, 22.44 tensile strength, and 118.41% elongation ratio, and they were higher than in the other two ADMs. There was no significant discrepancy in the findings of stiffness evaluation and suture retention strength test. The study had some limitations because there were many other more factors useful in ADM's testing. In the study, BellaCell HD showed complete decellularization, high biocompatibility, low cytotoxicity, high tensile strength, high elongation, and high suture retention strengths. These characteristics make BellaCell HD a suitable tissue for adequate and safe use in implant-based breast reconstruction in humans.

Biomechanical Properties and Biocompatibility of a Non-Absorbable Elastic Thread.

To date, extensive studies have been conducted to assess diverse types of sutures. But there is a paucity of data regarding biomechanical properties of commonly used suture materials. In the current experiment, we compared biomechanical properties and biocompatibility, such as tensile strength and elongation, the degree of bovine serum albumin (BSA) release, in vitro cytotoxicity and ex vivo frictional properties, between a non-absorbable elastic thread (NAT; HansBiomed Co. Ltd., Seoul, Korea) (NAT-R: NAT with a rough surface, NAT-S: NAT with a smooth surface) and the Elasticum® (Korpo SRL, Genova, Italy). The degree of tensile strength and elongation of Si threads was significantly higher in both the NAT-R and -S as compared with the Elasticum® (p < 0.05). Moreover, the degree of tensile strength and elongation of PET threads was significantly lower in both NAT-R and -S as compared with the Elasticum® (p < 0.05). Furthermore, the degree of tensile strength and elongation of braided Si/PET threads was significantly lower in NAT-S as compared with NAT-R and Elasticum® (p < 0.05). The degree of BSA release was significantly higher in the NAT-R as compared with Elasticum® and NAT-S throughout a 2-h period in the descending order (p < 0.05). The degree of cell viability was significantly higher in both NAT-R and -S as compared with Elasticum® (p < 0.05). The degree of coefficient of friction as well as the frictional force and strength was significantly higher in NAT-R as compared with NAT-S and Elasticum® (p < 0.05). NAT had a higher degree of biomechanical properties and biocompatibility as compared with Elasticum®. But further experimental and clinical studies are warranted to compare the efficacy, safety, and potential role as a carrier for drug delivery between NAT and Elasticum®.

Mechanical-chemical analyses and sub-chronic systemic toxicity of chemical treated organic bovine bone.

Sequentially chemical-treated bovine bone was not only evaluated by mechanical and chemical analyses but also implanted into the gluteal muscles of rats for 12 weeks to investigate potential local pathological effects and systemic toxicities. The test (chemical treated bone) and control (heat treated bone) materials were compared using scanning electron microscope (SEM), x-ray diffraction pattern, inductively coupled plasma analysis, and bending strength test. In the SEM images, the micro-porous structure of heat-treated bone was changed to sintered ceramic-like structure. The structure of bone mineral from test and control materials was analyzed as100% hydroxyapatite. The ratio of calcium (Ca) to potassium (P), the main inorganic elements, was same even though the Ca and P percentages of the control material was relatively higher than the test material. No death or critical symptoms arose from implantation of the test (chemical treated bone) and control (physiological saline) materials during 12 weeks. The implanted sites were macroscopically examined, with all the groups showing non-irritant results. Our results indicate that chemical processed bovine bone has a better mechanical property than the heat treated bone and the implantation of this material does not produce systemic or pathological toxicity.

Inactivation of enveloped and non-enveloped viruses in the process of chemical treatment and gamma irradiation of bovine-derived grafting materials.

BACKGROUND: Xenografts, unlike other grafting products, cannot be commercialized unless they conform to stringent safety regulations. Particularly with bovine-derived materials, it is essential to remove viruses and inactivate infectious factors because of the possibility that raw materials are imbrued with infectious viruses. The removal of the characteristics of infectious viruses from the bovine bone grafting materials need to be proved and inactivation process should satisfy the management provision of the Food and Drug Administration (FDA). To date, while most virus inactivation studies were performed in human allograft tissues, there have been almost no studies on bovine bone. METHODS: To evaluate the efficacy of virus inactivation after treatment of bovine bone with 70% ethanol, 4% sodium hydroxide, and gamma irradiation, we selected a variety of experimental model viruses that are known to be associated with bone pathogenesis, including bovine parvovirus (BPV), bovine herpes virus (BHV), bovine viral diarrhea virus (BVDV), and bovine parainfluenza-3 virus (BPIV-3). The cumulative virus log clearance factor or cumulative virus log reduction factor for the manufacturing process was obtained by calculating the sum of the individual virus log clearance factors or log reduction factors determined for individual process steps with different physicochemical methods. RESULTS: The cumulative log clearance factors achieved by three different virus inactivation processes were as follows: BPV ≥ 17.73, BHV ≥ 20.53, BVDV ≥ 19.00, and BPIV-3 ≥ 16.27. On the other hand, the cumulative log reduction factors achieved were as follows: BPV ≥ 16.95, BHV ≥ 20.22, BVDV ≥ 19.27, and BPIV-3 ≥ 15.58. CONCLUSIONS: Treatment with 70% ethanol, 4% sodium hydroxide, or gamma irradiation was found to be very effective in virus inactivation, since all viruses were at undetectable levels during each process. We have no doubt that application of this established process to bovine bone graft manufacture will be effective and essential.

Preparation of TGF-ß1-conjugated biodegradable pluronic F127 hydrogel and its application with adipose-derived stem cells.

In this study, a composite hydrogel using Pluronic F127 derivatives and crosslinked hyaluronic acid (X-HA) was investigated, exploring the benefits in the induction of chondrogenic differentiation of human adipose-derived stem cells (ASCs). F127 was chemically modified through a series of reactions that produced multiple F127 derivatives. A chondrogenic growth factor, transforming growth factor-beta 1 (TGF-ß1) was then coupled to the heparin-conjugated F127. X-HA was used as a physical stabilizer of the composite hydrogel (X-HA/F127). The chemical structures of F127 derivatives were analyzed using (1)H-NMR and ATR-FTIR. Sol-gel transition of the composite hydrogel was identified at body temperature. The conjugated TGF-ß1 was moderately released in vitro from the composite hydrogel. Cell viability of human ASCs in the hydrogels was about 50% after in vitro culture for 3 days. As the ASCs/hydrogel were injected into nude mice subcutaneously, DAPI staining of the retrieved constructs showed that ASCs were dispersed through the hydrogel matrix. From the immunofluorescent staining of type II collagen, the TGF-conjugated group exhibited more active green signals than the others. In addition, when those constructs were loaded into the full-thickness defect of rabbit knee articular cartilage, Alcian blue staining identified the formation of cartilaginous matrix from the TGF-conjugated hydrogel. The present work indicated that X-HA/F127 composite hydrogel was thermoreversible and biodegradable, and that the TGF-conjugated hydrogel could be effective in inducing chondrogenesis of human ASCs.