Retrospective comparison of postoperative dressing after eschar dermabrasion on paediatric scald wounds: Bacterial cellulose dressing and allogenic skin.

Eschar dermabrasion is an easy, cost-effective and dependable technique for debriding deep partial-thickness burn wounds, highly suitable for paediatric scalds. Postoperative dressing plays a crucial role in the subsequent healing process. While allogenic skin (AGS) has long been considered as the optimal coverage for abraded burn wounds by Chinese burn specialists, its clinical application on children has encountered challenges. In recent years, our department has observed promising results in the application of bacterial cellulose dressing on paediatric burn wounds after dermabrasion surgery. This study aimed to retrospectively review qualified cases from the past 5 years and categorize them into two groups: 201 cases in the AGS group and 116 cases in the bacterial cellulose dressing (BCD) group. Upon statistical analysis, no differences were oberved between the groups in terms of demographic information and wound characteristics. However, the BCD group had a significantly longer surgery time (44.3 ± 7.0 min vs. 31.5 ± 6.1 min, p < 0.01) and shorter healing time (19.6 ± 2.2 days vs. 24.4 ± 4.3 days, p < 0.01) compared to the AGS group. Moreover, the BCD group required fewer dressing changes (3.5 ± 0.8 vs. 6.7 ± 2.1, p < 0.01) and demonstrated lower rates of skin grafting (10/116 vs. 46/201, p = 0.036). In conclusion, our findings suggest that the bacterial cellulose material may serve as an optimal coverage option for paediatric abraded scald wounds.

Candidatus Accumulibacter use fermentation products for enhanced biological phosphorus removal.

Previous research suggested that two major groups of polyphosphate-accumulating organisms (PAOs), i.e., Ca. Accumulibacter and Tetrasphaera, play cooperative roles in enhanced biological phosphorus removal (EBPR). The fermentation of complex organic compounds by Tetrasphaera provides carbon sources for Ca. Accumulibacter. However, the viability of the fermentation products (e.g., lactate, succinate, alanine) as carbon sources for Ca. Accumulibacter and their potential effects on the metabolism of Ca. Accumulibacter were largely unknown. This work for the first time investigated the capability and metabolic details of Ca. Accumulibacter cognatus clade IIC strain SCUT-2 (enriched in a lab-scale reactor with a relative abundance of 42.8%) in using these fermentation products for EBPR. The enrichment culture was able to assimilate lactate and succinate with the anaerobic P release to carbon uptake ratios of 0.28 and 0.36 P mol/C mol, respectively. In the co-presence of acetate, the uptake of lactate was strongly inhibited, since two substrates shared the same transporter as suggested by the carbon uptake bioenergetic analysis. When acetate and succinate were fed at the same time, Ca. Accumulibacter assimilated two carbon sources simultaneously. Proton motive force (PMF) was the key driving force (up to 90%) for the uptake of lactate and succinate by Ca. Accumulibacter. Apart from the efflux of proton in symport with phosphate via the inorganic phosphate transport system, translocation of proton via the activity of fumarate reductase contributed to the generation of PMF, which agreed with the fact that PHV was a major component of PHA when lactate and succinate were used as carbon sources, involving the succinate-propionate pathway. Metabolic models for the usage of lactate and succinate by Ca. Accumulibacter for EBPR were built based on the combined physiological, biochemical, metagenomic, and metatranscriptomic analyses. Alanine was shown as an invalid carbon source for Ca. Accumulibacter. Instead, it significantly and adversely affected Ca. Accumulibacter-mediated EBPR. Phosphate release was observed without alanine uptake. Significant inhibitions on the aerobic phosphate uptake was also evident. Overall, this study suggested that there might not be a simply synergic relationship between Ca. Accumulibacter and Tetrasphaera. Their interactions would largely be determined by the kind of fermentation products released by the latter.

Using network pharmacology to fabricate crosslinked hyaluronan-chondroitin sulphate-resveratrol composite hydrogels for cartilage regeneration.

Cartilage tissue engineering provides a new approach for the treatment of cartilage damage. The combination of drug system with a tissue scaffold could be highly beneficial. Resveratrol (RES) is a potent anti-inflammatory agent, but its target genes and molecular mechanism of cartilage repair remain to be further studied. We used systems biology and network pharmacology methods to explore the mechanism of RES for chondrocyte and macrophages. Meanwhile, crosslinked hyaluronan-chondroitin sulphate-RES hydrogels (cHA-CS-RES) were constructed based on the target prediction results. Byin vitroandin vivoexperiments, we investigated its anti-inflammatory and pro-chondrogenesis. The results showed there were 12 hub genes potentially interacting in the RES-chondrocyte-macrophage network.In vitroexperiments were used to further verify the validity of the predicted hub genes. The composite hydrogels were successfully fabricated, and maintenance of the characteristic was further confirmed.In vitrostudy, cHA-CS-RES showed high cell viability, anti-inflammatory and pro-chondrogenesis abilities.In vivostudy of cartilage defects confirmed that the cHA-CS-RES groups were significantly better than the control group. Network pharmacology was used to predict and screen the target proteins of RES critical to cartilage tissue engineering. Moreover, cHA-CS-RES composite hydrogel showed good cartilage repair effects, anti-inflammatory and pro-chondrogenesis abilities.

Decryption for nitrogen removal in Anammox-based coupled systems: Nitrite-induced mechanisms.

This study investigated the effects of NO2- on synergetic interactions between Anammox bacteria (AnAOB) and sulfur-oxidizing bacteria (SOB) in an autotrophic denitrification-Anammox system. The presence of NO2- (0-75 mg-N/L) was shown to significantly enhance NH4+ and NO3- conversion rates, achieving intensified synergy between AnAOB and SOB. However, once NO2- exceed a threshold concentration (100 mg-N/L), both NH4+ and NO3- conversion rates decreased with increased NO2- consumption via autotrophic denitrification. The cooperation between AnAOB and SOB was decoupled due to the NO2- inhibition. Improved system reliability and nitrogen removal performance was achieved in a long-term reactor operation with NO2- in the influent; reverse transcription-quantitative polymerase chain reaction analysis showed elevated hydrazine synthase gene transcription levels (5.00-fold), comparing to these in the reactor without NO2-. This study elucidated the mechanism of NO2- induced synergetic interactions between AnAOB and SOB, providing theoretical guidance for engineering applications of Anammox-based coupled systems.

Simultaneous partial nitrification, Anammox and nitrate-dependent Fe(II) oxidation (NDFO) for total nitrogen removal under limited dissolved oxygen and completely autotrophic conditions.

Sustainable nitrogen removal from wastewater at reduced energy and/or chemical consumptions is challenging. This paper investigated, for the first time, the feasibility of coupled partial nitrification, Anammox and nitrate-dependent Fe(II) oxidation (NDFO) for sustainable autotrophic nitrogen removal. With NH4+-N as the only nitrogen-containing compound in the influent, near-complete nitrogen removal (a total of 97.5 % with a maximal total nitrogen removal rate of 6.64 ± 2.68 mgN/L/d) was achieved in a sequencing batch reactor for a 203-d operation without organic carbon source addition and forced aeration. Anammox (predominated by Candidatus Brocadia) and NDFO bacteria (such as Denitratisoma) were successfully enriched, with total relative abundances up to 11.54 % and 10.19 %, respectively. Dissolved oxygen (DO) concentration was a key factor affecting the coupling of multi (ammonia oxidization, Anammox, NDFO, iron-reduction, etc.) bacterial communities, resulting in different total nitrogen removal efficiencies and rates. In batch tests, the optimal DO concentration was 0.50-0.68 mg/L with a maximal total nitrogen removal efficiency of 98.7 %. Fe(II) in the sludge not only competed with nitrite oxidizing bacteria for DO to prevent complete nitrification, but promoted the transcription of NarG and NirK genes (10.5 and 3.5 times higher than the group without Fe(II) addition) as indicated by the reverse transcription quantitative polymerase chain reaction (RT-qPCR), resulting in increased NDFO rate (by 2.7 times) and promoted NO2--N generated from NO3--N, which back fed the Anammox process, achieving near-complete nitrogen removal. The reduction of Fe(III) by iron-reducing bacteria (IRB) and hydrolytic and fermentative anaerobes enabled a sustainable Fe(II)/Fe(III) recycling, avoiding the need in continuous Fe(II) or Fe (III) dosage. The coupled system is expected to benefit the development of novel autotrophic nitrogen removal processes with neglectable energy and material consumptions for the treatment of wastewater with low organic carbon and NH4+-N contents in underdeveloped regions, such as decentralized rural wastewaters.

Foliar methyl jasmonate (MeJA) application increased 2-acetyl-1-Pyrroline (2-AP) content and modulated antioxidant attributes and yield formation in fragrant rice.

Endogenous methyl jasmonate (MeJA) mediates abiotic and biotic stresses in plants. Exogenous MeJA application can stimulate and defend plant gene expression and induce plant chemical defense. The effects of foliar MeJA application on yield and 2-acetyl-1-pyrroline (2-AP) biosynthesis of fragrant rice are scarcely investigated. The pot experiment was conducted by spraying different concentrations of MeJA (0, 1, and 2 µM; denoted as CK, MeJA-1, and MeJA-2) at the initial heading stage of two fragrant rice cultivars, Meixiangzhan and Yuxiangyouzhan. The results showed that foliar MeJA application significantly increased the grain 2-AP content by 32.1% and 49.7%, respectively, following MeJA-1 and MeJA-2 treatments, and the two cultivars showed the highest 2-AP content upon MeJA-2 treatment. However, the grain yield was increased in MeJA-1 as compared with MeJA-2 treatment for all rice cultivars and no significant differences were observed in yield and yield-related traits compared with CK. The aroma was improved by foliar MeJA application which was strongly associated with the regulation of the precursors and enzymes involved in 2-AP biosynthesis. In particular, the contents of proline, pyrroline-5-carboxylic acid, and pyrroline at maturity, as well as the activities of proline dehydrogenase, ornithine aminotransferase, and pyrroline-5-carboxylic acid synthetase, were positively correlated with grain 2-AP content. On the other hand, foliar MeJA application improved the contents of soluble protein, chlorophyll a and b, and carotenoid, and increased the activity of antioxidant enzymes. Moreover, peroxidase activity and leaf chlorophyll contents were significantly positively correlated to 2-AP content following foliar MeJA application. Therefore, our results implied that foliar MeJA application increased aroma and influenced yield by regulating the physio-biochemistry characters and resistance, and suggested that the optimal concentration of MeJA for the best positive effect on the yield and aroma was 1 µM. However, further study is required to evaluate the metabolic level and molecular basis of the regulatory mechanism of foliar MeJA application on 2-AP in fragrant rice.

Identification of key biomarkers related to epithelial-mesenchymal transition and immune infiltration in ameloblastoma using integrated bioinformatics analysis.

OBJECTIVE: This study aimed to elucidate the underlying mechanisms of ameloblastoma (AM) through integrated bioinformatics analysis. METHODS: We downloaded two microarrays of AMs from the GEO database and identified differentially expressed genes (DEGs) by integrated bioinformatics analysis. The enrichment analysis of DEGs was conducted to characterize GO and KEGG pathways. Protein-protein interaction (PPI) network and hub genes were screened via STRING and Cytoscape. CIBERSORT algorithm was utilized to analyze immune infiltration in AMs. We also verified the diagnostic and therapeutic value of hub genes. RESULTS: Overall, 776 DEGs were identified in AMs through bioinformatics analysis. The function enrichment analysis shed light on pathways involved in AMs. Subsequently, we screened six hub genes via PPI network. Furthermore, we evaluated immune infiltration in AMs and found that macrophages may be participating in the progression of AMs. The upregulated expression of FN1 was related to the macrophages M2 polarization. Finally, ROC analysis indicated that six hub genes had high diagnostic value for AMs and 11 drugs interacted with upregulated hub genes were identified by screening the DGIdb database. CONCLUSION: This study revealed the underlying mechanisms of pathogenesis and biological behavior of AMs and provided candidate targets for the diagnosis and treatment of AMs.

Effect of electrohydrodynamic printing scaffold with different spacing on chondrocyte dedifferentiation.

Background: Osteoarthritis (OA) is a common degenerative disease. Chondrocyte dedifferentiation can accelerate the progress of OA. Three-dimensional printing (3DP) is widely used in tissue regeneration applications. A three-dimensional (3D) culture system with 3D printed scaffolds could reduce the dedifferentiation of chondrocytes during passages, which would be a potential method for chondrocyte expansion. Methods: The viability and proliferation of chondrocytes on scaffolds and effects of scaffolds with 100, 150, 200, 250 or 300 µm spacing on chondrocyte dedifferentiation were analyzed in vitro. The morphology of scaffolds and cell/scaffold constructs was observed by scanning electron microscopy (SEM). Glycosaminoglycan (GAG) was evaluated by Alcian blue staining. The effects of different spacing on chondrocyte dedifferentiation were evaluated by the messenger RNA (mRNA) and protein levels of cartilage-related genes. Results: With more binding sites, the proliferation and viability of chondrocytes on scaffolds with 100 and 150 µm spacing were better than those with 200, 250 and 300 µm spacing on day 1, but this advantage diminished over time. The histology and quantitative real-time polymerase chain reaction (qRT-PCR) results showed that 200 µm spacing inhibits chondrocyte dedifferentiation better. Conclusions: 3D printed scaffolds with 200 µm spacing can inhibit chondrocyte dedifferentiation, providing a basis for the future study of 3D printed scaffolds as an effective method for chondrocyte expansion.

ColoriSens: An open-source and low-cost portable color sensor board for microfluidic integration with wireless communication and fluorescence detection.

Microfluidic colorimetric biosensors have shown promising potential for detecting metal cations, anions, organic dyes, drugs, pesticides. As for today, most colorimetric sensors are read by a smartphone or professional optical imaging system, and there is still a lack of an affordable and reliable colorimetric detector for the microfluidic chip. Integrating those reading and detection capabilities into a microfluidic system is essential for point-of-care (POC) detection and can enable more complex microfluidic operations, such as lab-on-a-chip experiments or programmable microfluidics. We developed an open-source colorimetric detection sensor board that can be integrated into the existing microfluidic system. This sensor board has a built-in UV source that enables fluorescence detection. With built-in USB and Wi-Fi connectivity and a set of simple APIs, microfluidic systems can communicate directly with this sensor board, even wirelessly. The sensor was designed for low-cost. With a total build cost of less than 12 EUR per unit, it is ideal for low-cost systems and DIY microfluidic users. Along with the sensor board, we also designed a companion microfluidic chip carrier cartridge which can be modified depending on the chip's dimension. To demonstrate the sensor, we also developed a cross-platform open-source client application to demonstrate the communication APIs and the functionality of the sensor board.

Clinical application of 3D-printed patient-specific guide plate combined with computer navigation in acetabular reconstruction following resection of periacetabular tumors.

Background: The precise acetabular reconstruction has historically been a challenging procedure. 3D-printed patient-specific guide (PSG) and computer navigation (CN) technologies have been used to assist acetabular component positioning and pelvic reconstruction. This precise reconstruction approach may translate into clinical benefit. Methods: The clinical data of 84 patients who underwent periacetabular malignant tumor resection and screw-rod-acetabular cage system reconstruction in our center from January 2013 to December 2020 were retrospectively analyzed. Patients were divided into four groups: free hand (FH) group, PSG group, CN group, and PSG combined with computer navigation (PSG + CN) group. The operation time, intraoperative blood loss, and number of fluoroscopy views were recorded. The oncological prognosis, radiographic measurements of the acetabulum, limb function data, and postoperative complications were compared among groups. And finally, we evaluated the risk factors for mechanical failure of the prosthesis. Results: The postoperative X-ray and computed tomography (CT) scan revealed that the vertical offset discrepancy (VOD) between affected side and contralateral side was 8.4±1.9, 5.9±2.2, 4.1±1.3, and 2.4±1.2 mm in each groups; the horizontal offset discrepancy (HOD) was 9.0±1.9, 6.1±2.2, 3.2±1.3, and 2.1±1.2 mm, correspondingly; the abduction angle discrepancy (ABAD) was 8.6°±1.8°, 5.6°±2.0°, 2.5°±1.3°, and 1.8°±0.9°, respectively; the anteversion angle discrepancy (ANAD) was 5.9°±1.6°, 3.6°±1.7°, 2.9°±1.6°, and 1.9°±0.9°, correspondingly. Statistical results show that the PSG + CN group was superior to the FH group and the PSG group in terms of acetabular position and limb function (P<0.05). Body mass index (P=0.040) and resection type (P=0.042) were found to be the high-risk factors for mechanical failure of the prosthesis. Conclusions: PSG + CN has potential advantages in improving the accuracy and safety of acetabular positioning and reconstruction.

Carbon uptake bioenergetics of PAOs and GAOs in full-scale enhanced biological phosphorus removal systems.

This work analyzed, for the first time, the bioenergetics of PAOs and GAOs in full-scale wastewater treatment plants (WWTPs) for the uptake of different carbon sources. Fifteen samples were collected from five full-scale WWTPs. Predominance of different PAOs, i.e., Ca. Accumulibacter (0.00-0.49%), Tetrasphaera (0.37-3.94%), Microlunatus phosphovorus (0.01-0.18%), etc., and GAOs, i.e., Ca. Competibacter (0.08-5.39%), Defluviicoccus (0.05-5.34%), Micropruina (0.17-1.87%), etc., were shown by 16S rRNA gene amplicon sequencing. Despite the distinct PAO/GAO community compositions in different samples, proton motive force (PMF) was found as the key driving force (up to 90.1%) for the uptake of volatile fatty acids (VFAs, acetate and propionate) and amino acids (glutamate and aspartate) by both GAOs and PAOs at the community level, contrasting the previous understanding that Defluviicoccus have a low demand of PMF for acetate uptake. For the uptake of acetate or propionate, PAOs rarely activated F1, F0- ATPase (< 11.7%) or fumarate reductase (< 5.3%) for PMF generation; whereas, intensive involvements of these two pathways (up to 49.2% and 61.0%, respectively) were observed for GAOs, highlighting a major and community-level difference in their VFA uptake biogenetics in full-scale systems. However, different from VFAs, the uptake of glutamate and aspartate by both PAOs and GAOs commonly involved fumarate reductase and F1, F0-ATPase activities. Apart from these major and community-level differences, high level fine-scale micro-diversity in carbon uptake bioenergetics was observed within PAO and GAO lineages, probably resulting from their versatilities in employing different pathways for reducing power generation. Ca. Accumulibacter and Halomonas seemed to show higher dependency on the reverse operation of F1, F0-ATPase than other PAOs, likely due to the low involvement of glyoxylate shunt pathway. Unlike Tetrasphaera, but similar to Ca. Accumulibacter, Microlunatus phosphovorus took up glutamate and aspartate via the proton/glutamate-aspartate symporter driven by PMF. This feature was testified using a pure culture of Microlunatus phosphovorus stain NM-1. The major difference between PAOs and GAOs highlights the potential to selectively suppress GAOs for community regulation in EBPR systems. The finer-scale carbon uptake bioenergetics of PAOs or GAOs from different lineages benefits in understanding their interactions in community assembly in complex environment.

Portable all-in-one automated microfluidic system (PAMICON) with 3D-printed chip using novel fluid control mechanism.

State-of-the-art microfluidic systems rely on relatively expensive and bulky off-chip infrastructures. The core of a system-the microfluidic chip-requires a clean room and dedicated skills to be fabricated. Thus, state-of-the-art microfluidic systems are barely accessible, especially for the do-it-yourself (DIY) community or enthusiasts. Recent emerging technology-3D-printing-has shown promise to fabricate microfluidic chips more simply, but the resulting chip is mainly hardened and single-layered and can hardly replace the state-of-the-art Polydimethylsiloxane (PDMS) chip. There exists no convenient fluidic control mechanism yet suitable for the hardened single-layered chip, and particularly, the hardened single-layered chip cannot replicate the pneumatic valve-an essential actuator for automatically controlled microfluidics. Instead, 3D-printable non-pneumatic or manually actuated valve designs are reported, but their application is limited. Here, we present a low-cost accessible all-in-one portable microfluidic system, which uses an easy-to-print single-layered 3D-printed microfluidic chip along with a novel active control mechanism for fluids to enable more applications. This active control mechanism is based on air or gas interception and can, e.g., block, direct, and transport fluid. As a demonstration, we show the system can automatically control the fluid in microfluidic chips, which we designed and printed with a consumer-grade 3D-printer. The system is comparably compact and can automatically perform user-programmed experiments. All operations can be done directly on the system with no additional host device required. This work could support the spread of low budget accessible microfluidic systems as portable, usable on-the-go devices and increase the application field of 3D-printed microfluidic devices.

Investigating the protective effect of tanshinone IIA against chondrocyte dedifferentiation: a combined molecular biology and network pharmacology approach.

BACKGROUND: Osteoarthritis (OA) is a common degenerative disease with multifactorial etiology. The dedifferentiation of chondrocytes can accelerate the progress of OA. Tanshinone IIA (TIIA) has been widely used to treat OA for many years and has proved to be effective in inhibiting chondrocyte dedifferentiation. Until now, the precise mechanism of TIIA's effect against dedifferentiation has not been well understood. METHODS: The targets of TIIA were explored from public databases using various methods. The related targets of OA were obtained from the GeneCards database and the Online Mendelian Inheritance in Man (OMIM) database. The potential targets and signaling pathways were determined using protein-protein interaction (PPI), Gene Ontology (GO), and the Kyoto Encyclopedia of Genes and Genomes (KEGG). Cell viability, proliferation, and metabolic activity were analyzed in vitro. The effects of TIIA on chondrocyte dedifferentiation were evaluated by assessing morphological changes, glycosaminoglycan (GAG) production, and messenger RNA (mRNA) levels of cartilage-related genes. After 48 hours of culture in medium with 100 µg/mL TIIA, chondrocytes/hydrogel spheres were implanted to repair cartilage defects in a rat model. The harvested specimens were examined with hematoxylin and eosin (H&E) staining and immunohistochemistry to evaluate cartilage regeneration. RESULTS: The results showed that there were 28 genes potentially interacting in the TIIA-chondrocyte dedifferentiation network, and nine hub genes were identified. In vitro experiments showed an inhibitory effect of TIIA on chondrocyte dedifferentiation. The proliferation and viability of chondrocytes were promoted by TIIA at a concentration of 100-200 µg/mL, but inhibited by TIIA at 400 µg/mL. Furthermore, the histology results showed that chondrocyte/hydrogel spheres pre-treated with TIIA had better cartilage repair. CONCLUSIONS: This study revealed a systematic network pharmacology approach and provided a basis for the future study of TIIA as an effective treatment for cartilage regeneration. Moreover, in vitro and in vivo results confirmed the protective effects of TIIA against chondrocyte dedifferentiation.

Preoperative Axillary Ultrasound Helps in the Identification of a Limited Nodal Burden in Breast Cancer Patients.

Since the Z0011 trial, the clinical evaluation of axillary status has been redirected to predicting nodal tumor burden rather than nodal metastases. Our study aimed to evaluate the value of clinicopathological factors and axillary ultrasound (US) for the prediction of a high nodal burden (≥3 metastatic lymph nodes) in breast cancer patients. A total of 532 consecutive patients who underwent preoperative axillary US and subsequent surgery for clinical T1-2 breast cancer with a final pathologic analysis were included. Clinical and pathologic variables were retrospectively evaluated. Univariate and multivariate statistical analyses were performed to identify the variables that were associated with a high nodal burden. Among the 532 patients, 110 (20.7%) had a high axillary nodal burden and 422 (79.3%) had a limited nodal burden. The multivariate analysis showed that suspicious axillary US findings (P < 0.001), clinical T2 stage (P = 0.011), the presence of lymphovascular invasion (P < 0.001), and estrogen receptor positivity (P < 0.001) were significantly associated with a high nodal burden. Patients with negative axillary US findings seldom had a high nodal burden, with a negative predictive value of 93.0% (294/316). Patients with suspicious axillary US findings, clinical T2 stage, lymphovascular invasion, and estrogen receptor positivity are more likely to have a high nodal burden, which may provide additional information for the treatment plan of breast cancer patients. Preoperative axillary US helps identify a limited nodal burden in breast cancer patients and has implications for axillary lymph node dissection and adjuvant treatment.

Implementation of a stratified approach and gene immobilization to enhance the osseointegration of a silk-based ligament graft.

A silk scaffold exhibits high potential for the human anterior cruciate ligament (ACL) reconstruction due to its exceptional mechanics as well as biocompatibility. Inefficient ACL interface restoration is thought to be a major hurdle for the common implementation of a silk-based ligament graft. By integrating a stratified approach and gene immobilization, here we developed a gene-immobilized triphasic silk scaffold to enhance ACL osseointegration. Isotropic silk was divided into three regions (respectively corresponding to a ligament, fibrocartilage and the bone region of the native ACL interface) using a custom-made divider, and the lentiviral vector-encoded transforming growth factor beta-3 (TGF-ß3) and bone morphogenetic protein-2 (BMP2) was further, respectively, immobilized to phosphatidylserine (PS)-coated fibrocartilage and the bone region of the triphasic silk scaffold. The in vitro assessments displayed that this gene-immobilized triphasic silk scaffold significantly promotes bone marrow mesenchymal stem cell (BMSC) proliferation and differentiation into corresponding cell lineage. Moreover, the gene-modified triphasic silk scaffold combined with BMSCs alone, which was rolled into a compact shaft to be implanted onto rabbit ACL-defect models, revealed roughly complete osseointegration restoration as a result of apparent three-layered tissue formation and robust mechanical ability as early as 12 weeks postoperatively. These outcomes demonstrated that employing the stratified approach and gene immobilization efficiently expedites silk-mediated ACL interface formation, expanding the therapeutic potential of the silk-based ligament graft for ACL reconstruction.