Decellularization of human dermis using non-denaturing anionic detergent and endonuclease: a review

Abstract

Decellularized human dermis has been used for a number of clinical applications including wound healing, soft tissue reconstruction, and sports medicine procedures. A variety of methods exist to prepare this useful class of biomaterial. Here, we describe a decellularization technology (MatrACELL(®)) utilizing a non-denaturing anionic detergent, N-Lauroyl sarcosinate, and endonuclease, which was developed to remove potentially immunogenic material while retaining biomechanical properties. Effective decellularization was demonstrated by a residual DNA content of ≤4 ng/mg of wet weight which represented >97 % DNA removal compared to unprocessed dermis. Two millimeter thick MatrACELL processed human acellular dermal matrix (MH-ADM) exhibited average ultimate tensile load to failure of 635.4 ± 199.9 N and average suture retention strength of 134.9 ± 55.1 N. Using an in vivo mouse skin excisional model, MH-ADM was shown to be biocompatible and capable of supporting cellular and vascular in-growth. Finally, clinical studies of MH-ADM in variety of applications suggest it can be an appropriate scaffold for wound healing, soft tissue reconstruction, and soft tissue augmentation.

Fig. 1

Histological analysis of human skin tissue. a prior to and b after MatrACELL processing. Hemotoxylin and eosin (H&E) staining shows nuclear material (blue/purple staining). Note the presence of stained nuclear material prior to decellularization (a) in contrast to the lack of nuclear material in the tissue after the MatrACELL process in MH-ADM (b). (Data on file at LifeNet Health). (Color figure online)

Fig. 2

Major Histocompatibility Complex I (MHC-I) staining of human skin tissue a prior to and b after the MatrACELL process. MHC-I staining was used to detect cells and cellular remnants. Note the presence of brick-red stained MHC-I positive cells prior to decellularization (a) in contrast to the absence of MHC-I staining in the tissue after the MatrACELL process (b). (Data on file at LifeNet Health). (Color figure online)

Fig. 3

Comparison of residual DNA content in three ADM. All DNA content results are presented as ng/mg of dry weight of material. Please note this is not a side-by-side experiment, rather a comparison to the literature. However, all values are represented as ng/mg dry weight of tissue, and the method of detection was identical. Asterisk MH-ADM (Data on file at LifeNet Health, n = 3 as 1 sample each from 3 donors) and values reported in the literature for: ^ GraftJacket (Derwin et al. 2010), ⁿ AlloDerm (Choe and Bell 2001)

Fig. 4

Suture pullout strength comparison of two thickness MH-ADM (2 mm* and 1.5 mm*) compared with two thickness GRAFTJACKET and other surgical mesh products (^). Data generated for MatrACELL Dermis and all other materials, respectively was generated at different points in time; however, the exact same methods, fixtures, material testing machine, and facility was used for both studies. Asterisk Data on File at Arthrex (n = 5), ^ Barber and Aziz-Jacobo (2009)

Fig. 5

Ultimate load to failure comparison of two thickness MH-ADM (2 and 1.5 mm*) is compared with two thickness GRAFTJACKET and other surgical mesh products (^). Data generated for MatrACELL Dermis and all other materials, respectively was generated at different points in time; however, the exact same methods, fixtures, material testing machine, and facility was used for both studies. Asterisk Data on File at Arthrex, ^ Barber and Aziz-Jacobo (2009)

Fig. 6

Hematoxylin and eosin staining of MH-ADM explants using nude mouse skin excisional model. The implant was in place for 16 days prior to excision and analysis. Note the presence of new blood vessels (black arrows) and epithelial layer (open arrow) (data on file at LifeNet Health). (Color figure online)

Fig. 7

Use of MH-ADM to augment an Achilles tendon repair. Photo courtesy of Jeffrey Barton, DPM (Delaware Surgery Center, Dover, DE) showed before (a) and after (b) the augmentation

Fig. 8

Use of MH-ADM to augment a biceps tendon repair. Photo courtesy of Raffy Mirzayan, MD, Los Angeles, CA

Fig. 9

Use of MH-ADM in conjunction with cortical bone particulates to correct for the thin bone implant support and to increase the soft tissue profile in these areas. a Pre-surgery showing a thin bone ridge. b The application of MH-ADM in conjunction with implant bone and pins. c 4 weeks post-implant demonstrating increase in tissue profile and a smoothly healed gum line. The case photos are courtesy of Paul Rosen, DMD, MS (Yardley, PA)

Fig. 10

Use of MH-ADM to repair a temporal depression. a Pre-surgery showing the temporal depression. b The depression smoothing following an incision in the hair line and insertion of MH-ADM. The case photos are courtesy of Barry Eppley, MD (Indianapolis, IN)

Fig. 11

Use of MH-ADM for wound repair of a diabetic foot ulcer. a 47 y.o. female presented with a Wagner Grade 2 non-healing diabetic ulcer on the plantar first medial head. b The patient went on to a successful outcome with a single application of MH-ADM as noted by the substantial healing at week 12 (Yonehiro et al. : reproduced with permission of the authors)

Fig. 12

Use of MH-ADM for breast reconstruction. a pre-operative and b post-operative. In this case, a 46 y.o patient received a bilateral mastectomy and advanced to the 2nd stage repair. Her expanders were filled to the full 550 cc and replaced with a 700 cc silicone implant 16 weeks following mastectomy (Vashi : reproduced with permission of the author)

Fig. 13

Hematoxylin and eosin staining of biopsy specimen following 16 weeks in situ placement of MH-ADM in the breast reconstruction surgery shown in Fig. 12. Note the intact ultrastructure and also evidence of cellular in-growth with apparent fibroblasts (arrows) at 10× magnification (data on file at LifeNet Health). (Color figure online)