Silk fibroin and hydroxyapatite segmented coating enhances graft ligamentization and osseointegration processes of the polyethylene terephthalate artificial ligament in vitro and in vivo

Cai, Jiangyu; Wan, Fang; Dong, Qinglin; Jiang, Jia; Ai, Chengchong; Sheng, Dandan; Jin, Wenhe; Liu, Xingwang; Zhi, Yunlong; Wang, Siheng; Sun, Yaying; Shao, Zhengzhong; Chen, Shiyi

doi:10.1039/c8tb01310a

Cited by 23 publications

(21 citation statements)

References 27 publications

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“…A thinner interface suggests better integration. 4 For evaluating the maturity of the intra-articular portion of the graft (midsubstance), the content of collagen 1 (Col 1) by sirius red staining, as indicated by yellow-red strong birefringence, was viewed by polarized microscopy. 41 A light microscope was used to observe the a-SMA1 area.…”

Section: Histological Experimentsmentioning

confidence: 99%

Stem Cell–Conditioned Medium Promotes Graft Remodeling of Midsubstance and Intratunnel Incorporation After Anterior Cruciate Ligament Reconstruction in a Rat Model

et al. 2019

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Background: Stem cell–conditioned medium (CM) has been increasingly used in regenerative medicine. However, its effect on graft-host integration after anterior cruciate ligament (ACL) reconstruction (ACLR) remains unclear. Purpose: To examine the effect of human bone marrow stem cell (hBMSC)–CM on graft-bone integration and graft midsubstance ligamentization in a rat model of ACLR. Study Design: Controlled laboratory study. Methods: CM was obtained from the supernatant of commercially available hBMSCs in serum-free Dulbecco’s modified Eagle medium (DMEM). In a rat model of an ACL injury, isometric ACLR was performed. Three groups were established: CM injection group (CM; n = 40), control injection group (CI; n = 40) with serum-free DMEM injections, and no injection group (NI; n = 40). An intra-articular injection was performed weekly. Micro–computed tomography was conducted at 2, 4, and 8 weeks postoperatively. Histological and biomechanical analyses were conducted at 4 and 8 weeks postoperatively. The NIH3T3 fibroblast was utilized as a model in vitro to examine the effect of CM using the cell counting kit–8 (CCK-8) assay and immunofluorescence staining of Ki-67, α–smooth muscle actin (α-SMA), and collagen 1 (Col 1). Results: At 4 and 8 weeks, the femoral and tibial bone tunnel areas as well as the interface between the graft and host bone were smaller, while the bone volume/total volume ratio was higher, in the CM group. Sharpey-like fibers formed at 8 weeks in the CM group. At 4 and 8 weeks, more Col 1 was noticed in the CM group than in the NI group (both P < .001) or CI group (both P < .001). Immunohistochemically, the α-SMA–positive area was up-regulated at the graft-bone interface at 4 weeks ( P < .001) and declined at 8 weeks ( P < .001) in the CM group compared with the other 2 groups. At the midsubstance, α-SMA expression decreased from 4 to 8 weeks in all groups and was significantly lower in the CM group than in the NI group ( P < .01) or CI group ( P < .05) at 8 weeks. The CCK-8 assay showed that CM increased NIH3T3 viability ( P < .001) and the level of Ki-67 ( P < .05), α-SMA ( P < .001), and Col 1 ( P < .001) in CM-educated NIH3T3 cells. Conclusion: hBMSC-CM accelerates graft-bone incorporation and midsubstance ligamentization and enhances the proliferation, differentiation, and collagen synthesis of fibroblasts. Clinical Relevance: Graft-host integration is essential after ACLR. The current study identified a novel agent, that is, hBMSC-CM, as a candidate for promoting integration.

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Section: Histological Experimentsmentioning

confidence: 99%

Stem Cell–Conditioned Medium Promotes Graft Remodeling of Midsubstance and Intratunnel Incorporation After Anterior Cruciate Ligament Reconstruction in a Rat Model

et al. 2019

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“…A smaller interface width indicated better and more reliable tendon-bone healing. 7 The tendon-bone interface width was significantly narrower in the AT group than in the HB group from 4 to 12 weeks. These differences indicate that using an autograft for augmentation in ACL reconstruction may promote tendon-bone healing.…”

Section: Discussionmentioning

confidence: 84%

“…As noted in previous studies, a thinner interface suggests better integration. 4,7,8,47 The mean interface width of a sample was calculated by averaging the width at every 30° point of the tunnel cross section from 0° to 360°, and 3 sections were measured per sample. 47 The graft-bone integration parameters were analyzed using a previously validated scoring system.…”

Section: Methodsmentioning

confidence: 99%

An Autograft for Anterior Cruciate Ligament Reconstruction Results in Better Biomechanical Performance and Tendon-Bone Incorporation Than Does a Hybrid Graft in a Rat Model

Wang

Sui

et al. 2020

Am J Sports Med

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Background: The biomechanical and tendon-bone incorporation properties of allograft-augmented hybrid grafts for anterior cruciate ligament (ACL) reconstruction compared with traditional autografts are unknown. Hypothesis: Using an autograft for ACL reconstruction yields better results on biomechanical testing, radiographic analysis, and histological evaluation versus using a hybrid graft. Study Design: Controlled laboratory study. Methods: A total of 66 adult male Sprague Dawley rats underwent unilateral ACL reconstruction with an autograft (AT group; n = 33) or a hybrid graft (HB group; n = 33). The grafts used in both groups were harvested from the peroneus longus tendon and were fixed by suturing to the surrounding periosteum. Samples were harvested for biomechanical testing, micro–computed tomography (CT), and histological evaluation at 4, 8, and 12 weeks postoperatively. Bone tunnels on the femoral and tibial sides were divided into 3 subregions: intra-articular (IA), midtunnel (MT), and extra-articular (EA). A cylinder-like volume of interest in the bone tunnel and a tubular-like volume of interest around the bone tunnel were used to evaluate new bone formation and bone remodeling, respectively, via micro-CT. Results: In the AT group, there were significantly higher failure loads and stiffness at 8 weeks (failure load: 3.04 ± 0.40 vs 2.09 ± 0.54 N, respectively; P = .006) (stiffness: 3.43 ± 0.56 vs 1.75 ± 0.52 N/mm, respectively; P < .001) and 12 weeks (failure load: 9.10 ± 1.13 vs 7.14 ± 0.94 N, respectively; P = .008) (stiffness: 4.45 ± 0.75 vs 3.36 ± 0.29 N/mm, respectively; P = .008) than in the HB group. With regard to new bone formation in the bone tunnel, in the AT group, the bone volume/total volume (BV/TV) was significantly higher than in the HB group on the tibial side at 8 weeks (IA: 22.21 ± 4.98 vs 5.16 ± 3.98, respectively; P < .001) (EA: 19.66 ± 7.19 vs 10.85 ± 2.16, respectively; P = .030) and 12 weeks (IA: 30.50 ± 5.04 vs 17.11 ± 7.31, respectively; P = .010) (MT: 21.15 ± 2.58 vs 15.55 ± 4.48, respectively; P = .041) (EA: 20.75 ± 3.87 vs 10.64 ± 3.94, respectively; P = .003). With regard to bone remodeling around the tunnel, the BV/TV was also significantly higher on the tibial side at 8 weeks (MT: 33.17 ± 8.05 vs 15.21 ± 7.60, respectively; P = .007) (EA: 25.19 ± 6.38 vs 13.94 ± 7.10, respectively; P = .030) and 12 weeks (IA: 69.46 ± 4.45 vs 47.80 ± 6.16, respectively; P < .001) (MT: 33.15 ± 3.88 vs 13.76 ± 4.07, respectively; P < .001) in the AT group than in the HB group. Sharpey-like fibers had formed at 8 weeks in the AT group. A large number of fibroblasts withdrew at 12 weeks. In the AT group, the width of the interface was significantly narrower at 4 weeks (85.86 ± 17.49 vs 182.97 ± 14.35 μm, respectively; P < .001), 8 weeks (58.86 ± 10.99 vs 90.15 ± 11.53 μm, respectively; P = .002), and 12 weeks (42.70 ± 7.96 vs 67.29 ± 6.55 μm, respectively; P = .001) than in the HB group. Conclusion: Using an autograft for ACL reconstruction may result in improved biomechanical properties and tendon-bone incorporation compared with a hybrid graft. Clinical Relevance: Augmenting small autografts with allograft tissue may result in decreased biomechanical performance and worse tendon-bone incorporation, increasing the risk of graft failure.

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“…Using micro-CT and histological analysis, the current study comprehensively evaluated the effects of SV/CS-encapsulated PET-HA ligaments on the osteointegration process in a rat extra-articular model. Although the anterior cruciate ligament reconstruction model likely reflects the clinical situation, one fact should not be neglected: the complex microenvironments within the joint, including the biomechanical factors, as well as the complicated surgical factors, might result in poor reproducibility of the surgery outcome 37. Consequently, this insufficiency in consistency further magnifies the bias in the accuracy of the outcome.…”

Section: Resultsmentioning

confidence: 99%

Encapsulation of a nanoporous simvastatin-chitosan composite to enhance osteointegration of hydroxyapatite-coated polyethylene terephthalate ligaments

Ding

Wang

Jin

et al. 2019

IJN

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Purpose This study was designed to evaluate the in vitro and in vivo biocompatibility and osteointegration of plasma-sprayed hydroxyapatite (HA)-coated polyethylene terephthalate (PET) ligaments encapsulated with a simvastatin (SV)-chitosan (CS) composite. Methods This study compared the in vitro and in vivo bone responses to three different PET ligaments: SV/CS/PET-HA, CS/PET-HA and PET-HA. A field emission scanning electron microscope was used to characterize the morphology, and the in vitro SV release profile was analyzed. MC3T3 cells were cocultured with SV/CS/PET-HA, CS/PET-HA and PET-HA to test their biocompatibility using CCK-8 tests. Osteogenic differentiation was investigated by the expression of marker genes using qPCR. Osteointegration was performed by implanting the PET ligaments into the proximal tibia bone tunnels of male Sprague-Dawley rats for 3 weeks and 6 weeks. The bone-implant interface was evaluated by micro-computed tomography (micro-CT) and histological analysis. Results The characteristic nanoporous structures mainly formed on the surface of the plasma-sprayed HA particles in the SV/CS/PET-HA and CS/PET-HA groups. The SV release test showed that the sustained release of simvastatin lasted for 25 days in the SV/CS/PET-HA group. The in vitro studies demonstrated that the SV/CS/PET-HA ligaments induced osteogenic differentiation in the MC3T3 cells, with higher mRNA expression levels of collagen-1, bone morphogenetic protein-2, osteocalcin and alkaline phosphatase than those in the CS/PET-HA and PET-HA ligament groups. The in vivo tests showed that both micro-CT analysis (bone mineral density and bone volume per total volume) and histological analysis (bone implant contact and interface area) revealed significantly higher peri-implant bone formation and less interface area in the SV/CS/PET-HA group than in the other groups. Conclusion The SV-CS composite nanoporous structure was associated with the improved biocompatibility and osteogenic differentiation in vitro and enhanced osteointegration process in vivo of plasma-sprayed HA-coated PET ligaments.

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Silk fibroin and hydroxyapatite segmented coating enhances graft ligamentization and osseointegration processes of the polyethylene terephthalate artificial ligament in vitro and in vivo

Abstract: A silk fibroin and hydroxyapatite segmented coating ligament is fabricated to enhances graft ligamentization and osseointegration processes successfully.

Cited by 23 publications

References 27 publications

Stem Cell–Conditioned Medium Promotes Graft Remodeling of Midsubstance and Intratunnel Incorporation After Anterior Cruciate Ligament Reconstruction in a Rat Model

Stem Cell–Conditioned Medium Promotes Graft Remodeling of Midsubstance and Intratunnel Incorporation After Anterior Cruciate Ligament Reconstruction in a Rat Model

An Autograft for Anterior Cruciate Ligament Reconstruction Results in Better Biomechanical Performance and Tendon-Bone Incorporation Than Does a Hybrid Graft in a Rat Model

<p>Encapsulation of a nanoporous simvastatin-chitosan composite to enhance osteointegration of hydroxyapatite-coated polyethylene terephthalate ligaments</p>

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