Physicochemical decellularization of composite flexor tendon-bone interface grafts

Joel A. Bronstein, Colin Y L Woon, Simon Farnebo, Anthony W. Behn, Taliah Schmitt, Hung Pham, Alesha Castillo, James Chang

Research output: Contribution to journalArticle

Abstract

BACKGROUND: Extremity injuries involving tendon attachment to bone are difficult to address. Clinically, tendon-bone interface allografts must be decellularized to reduce immunogenicity. Composite grafts are difficult to decellularize because chemical agents cannot reach cells between tissues. In this study, the authors attempted to optimize tendon-bone interface graft decellularization. METHODS: Human flexor digitorum profundus tendons with attached distal phalanx were harvested from cadavers and divided into four groups. Group 1 (control) was untreated. Group 2 (chemical) was chemically treated with 5% peracetic acid, 0.1% ethylenediaminetetraacetic acid, and 0.1% sodium dodecyl sulfate. Group 3 (low-power) underwent targeted ultrasonication for 3 minutes (22,274 J, 126W) followed by chemical decellularization. Group 4 (high-power) underwent targeted ultrasonication for 10 minutes (88,490 J, 155W) followed by chemical decellularization. Decellularization was assessed histologically with hematoxylin and eosin stain and stains for major histocompatibility complex I stains. Cell counts were performed. The ultimate tensile load of decellularized grafts (group 4) were compared with pair-matched untreated grafts (group 1). RESULTS: Average cell counts were 100 ± 41, 27 ± 10, 12 ± 11, and 6 ± 11 per high-power field for groups 1, 2, 3, and 4, respectively (p < 0.001). Decellularization using physical and chemical treatments (groups 3 and 4) resulted in substantial reduction of cells and major histocompatibility complex I molecules. There was no difference in ultimate tensile load between treated (group4) and untreated (group 1) samples (p > 0.5). CONCLUSIONS: Physicochemical decellularization of tendon-bone interface grafts using targeted ultrasonication and chemical treatment resulted in near-complete reduction in cellularity and maintenance of tensile strength. In the future, these decellularized composite scaffolds may be used for reconstruction of tendon-bone injuries.

Original languageEnglish (US)
Pages (from-to)94-102
Number of pages9
JournalPlastic and Reconstructive Surgery
Volume132
Issue number1
DOIs
StatePublished - Jul 2013

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Tendons
Transplants
Bone and Bones
Tendon Injuries
Coloring Agents
Cell Count
Peracetic Acid
Tensile Strength
Hematoxylin
Eosine Yellowish-(YS)
Major Histocompatibility Complex
Cadaver
Edetic Acid
Sodium Dodecyl Sulfate
Allografts
Extremities
Maintenance
Control Groups

ASJC Scopus subject areas

  • Surgery

Cite this

Bronstein, J. A., Woon, C. Y. L., Farnebo, S., Behn, A. W., Schmitt, T., Pham, H., ... Chang, J. (2013). Physicochemical decellularization of composite flexor tendon-bone interface grafts. Plastic and Reconstructive Surgery, 132(1), 94-102. https://doi.org/10.1097/PRS.0b013e318290f5fc

Physicochemical decellularization of composite flexor tendon-bone interface grafts. / Bronstein, Joel A.; Woon, Colin Y L; Farnebo, Simon; Behn, Anthony W.; Schmitt, Taliah; Pham, Hung; Castillo, Alesha; Chang, James.

In: Plastic and Reconstructive Surgery, Vol. 132, No. 1, 07.2013, p. 94-102.

Research output: Contribution to journalArticle

Bronstein, JA, Woon, CYL, Farnebo, S, Behn, AW, Schmitt, T, Pham, H, Castillo, A & Chang, J 2013, 'Physicochemical decellularization of composite flexor tendon-bone interface grafts', Plastic and Reconstructive Surgery, vol. 132, no. 1, pp. 94-102. https://doi.org/10.1097/PRS.0b013e318290f5fc
Bronstein, Joel A. ; Woon, Colin Y L ; Farnebo, Simon ; Behn, Anthony W. ; Schmitt, Taliah ; Pham, Hung ; Castillo, Alesha ; Chang, James. / Physicochemical decellularization of composite flexor tendon-bone interface grafts. In: Plastic and Reconstructive Surgery. 2013 ; Vol. 132, No. 1. pp. 94-102.
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abstract = "BACKGROUND: Extremity injuries involving tendon attachment to bone are difficult to address. Clinically, tendon-bone interface allografts must be decellularized to reduce immunogenicity. Composite grafts are difficult to decellularize because chemical agents cannot reach cells between tissues. In this study, the authors attempted to optimize tendon-bone interface graft decellularization. METHODS: Human flexor digitorum profundus tendons with attached distal phalanx were harvested from cadavers and divided into four groups. Group 1 (control) was untreated. Group 2 (chemical) was chemically treated with 5{\%} peracetic acid, 0.1{\%} ethylenediaminetetraacetic acid, and 0.1{\%} sodium dodecyl sulfate. Group 3 (low-power) underwent targeted ultrasonication for 3 minutes (22,274 J, 126W) followed by chemical decellularization. Group 4 (high-power) underwent targeted ultrasonication for 10 minutes (88,490 J, 155W) followed by chemical decellularization. Decellularization was assessed histologically with hematoxylin and eosin stain and stains for major histocompatibility complex I stains. Cell counts were performed. The ultimate tensile load of decellularized grafts (group 4) were compared with pair-matched untreated grafts (group 1). RESULTS: Average cell counts were 100 ± 41, 27 ± 10, 12 ± 11, and 6 ± 11 per high-power field for groups 1, 2, 3, and 4, respectively (p < 0.001). Decellularization using physical and chemical treatments (groups 3 and 4) resulted in substantial reduction of cells and major histocompatibility complex I molecules. There was no difference in ultimate tensile load between treated (group4) and untreated (group 1) samples (p > 0.5). CONCLUSIONS: Physicochemical decellularization of tendon-bone interface grafts using targeted ultrasonication and chemical treatment resulted in near-complete reduction in cellularity and maintenance of tensile strength. In the future, these decellularized composite scaffolds may be used for reconstruction of tendon-bone injuries.",
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AU - Schmitt, Taliah

AU - Pham, Hung

AU - Castillo, Alesha

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AB - BACKGROUND: Extremity injuries involving tendon attachment to bone are difficult to address. Clinically, tendon-bone interface allografts must be decellularized to reduce immunogenicity. Composite grafts are difficult to decellularize because chemical agents cannot reach cells between tissues. In this study, the authors attempted to optimize tendon-bone interface graft decellularization. METHODS: Human flexor digitorum profundus tendons with attached distal phalanx were harvested from cadavers and divided into four groups. Group 1 (control) was untreated. Group 2 (chemical) was chemically treated with 5% peracetic acid, 0.1% ethylenediaminetetraacetic acid, and 0.1% sodium dodecyl sulfate. Group 3 (low-power) underwent targeted ultrasonication for 3 minutes (22,274 J, 126W) followed by chemical decellularization. Group 4 (high-power) underwent targeted ultrasonication for 10 minutes (88,490 J, 155W) followed by chemical decellularization. Decellularization was assessed histologically with hematoxylin and eosin stain and stains for major histocompatibility complex I stains. Cell counts were performed. The ultimate tensile load of decellularized grafts (group 4) were compared with pair-matched untreated grafts (group 1). RESULTS: Average cell counts were 100 ± 41, 27 ± 10, 12 ± 11, and 6 ± 11 per high-power field for groups 1, 2, 3, and 4, respectively (p < 0.001). Decellularization using physical and chemical treatments (groups 3 and 4) resulted in substantial reduction of cells and major histocompatibility complex I molecules. There was no difference in ultimate tensile load between treated (group4) and untreated (group 1) samples (p > 0.5). CONCLUSIONS: Physicochemical decellularization of tendon-bone interface grafts using targeted ultrasonication and chemical treatment resulted in near-complete reduction in cellularity and maintenance of tensile strength. In the future, these decellularized composite scaffolds may be used for reconstruction of tendon-bone injuries.

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