Intramuscular Implantation Research Articles

Autologous myokine-loaded pre-vascularized bioactive scaffold enhances bone augmentation

The poor angiogenic ability is the main reason for the failure of bone augmentation material implantation. Pre-vascularized culture is considered to be an effective method to accelerate early angiogenesis, while the immune rejection has limited clinical application. Herein, since bone augmentation is an elective procedure, an easily accessible pre-vascularized silk fibroin/bioactive glass (SF-BG) scaffold without immune rejection was prepared by autologous intramuscular implantation. The SF-BG scaffolds exhibited outstanding vascularization ability in muscle by enhancing the muscle endocrine function. Further mechanism study confirmed that BG improved the synthesis and secretion of myokine irisin by regulating PI3K/Akt/PGC-1α/FNDC5 signaling pathway. After implantation in the bone augmentation position, the pre-vascularized BG scaffold with irisin loaded fostered the early angiogenesis of implantation and increased bone augmentation at the late stage. This study proposed a new idea for bone augmentation by autologous intramuscular pre-vascularized scaffolds.

Incorporating Microbial Stimuli for Osteogenesis in a Rabbit Posterolateral Spinal Fusion Model.

Autologous bone grafts are commonly used to repair defects in skeletal tissue, however, due to their limited supply there is a clinical need for alternatives. Synthetic ceramics present a promising option but currently lack biological activity to stimulate bone regeneration. One potential approach to address this limitation is the incorporation of immunomodulatory agents. In this study, we investigate the application of microbial stimuli to stimulate bone formation. Three different microbial stimuli were incorporated in a biphasic calcium phosphate (BCP) ceramic: Bacille Calmette-Guérin (BCG), gamma-irradiated Staphylococcus aureus (γi-S. aureus), or Candida albicans (γi-C. Albicans). The constructs were then implanted in both a rabbit posterolateral spinal fusion (PLF) and an intramuscular implant model for 10 weeks and compared to a non-stimulated control construct. For the PLF model, the formation of a bony bridge was evaluated by manual palpation, micro-CT, and histology. While complete fusion was not observed, the BCG condition was most promising with higher manual stiffness and almost twice as much bone volume in the central fusion mass compared to the control (9±4.4% bone area vs 4.6±2.3%, respectively). Conversely, the γi-S. aureus or γi-C. albicans appeared to inhibit bone formation (1.4±1.4% and 1.2±0.6% bone area). Bone induction was not observed in any of the intramuscular implants. This study indicates that incorporating immunomodulatory agents in ceramic bone substitutes can affect bone formation, which can be positive when selected carefully. The readily available and clinically approved BCG showed promising results, which warrants further research for clinical translation.

Biocompatibility Study of Purified and Low-Temperature-Sterilized Injectable Collagen for Soft Tissue Repair: Intramuscular Implantation in Rats.

The clinical application of collagen-based biomaterials is expanding rapidly, especially in tissue engineering and cosmetics. While oral supplements and injectable skin boosters are popular for enhancing skin health, clinical evidence supporting their effectiveness remains limited. Injectable products show potential in revitalizing skin, but safety concerns persist due to challenges in sterilization and the risk of biological contamination. Traditional methods of sterilization (heat and irradiation) can denature collagen. This study addresses these issues by introducing a novel technique: the double filtration and low-temperature steam sterilization of a collagen gel. In vitro tests documented the sterility and confirmed that the collagen did not show cytotoxicity, degradation, integrity, and viscosity characteristics changes after the processing and sterilization. The collagen gel induced new collagen expression and the proliferation of human dermal fibroblasts when the cells were cultured with the collagen gel. An in vivo study found no adverse effects in rats or significant lesions at the implantation site over 13 weeks. These results suggest that this novel method to process collagen gels is a safe and effective skin booster. Advanced processing methods are likely to mitigate the safety risks associated with injectable collagen products, though further research is needed to validate their biological effectiveness and clinical benefits.

3D printing calcium phosphate ceramics with high osteoinductivity through pore architecture optimization

The osteoinductivity of 3D printed calcium phosphate (CaP) ceramics has a large gap compared with those prepared by conventional foaming methods, and improving the osteoinductivity of 3D printing CaP ceramics is crucial for successful application in bone regeneration. Pore architecture plays a critical role in osteoinductivity. In this study, CaP ceramics with a hexagonal close-packed (HCP) spherical pore structure were successfully fabricated using DLP printing technology. Additionally, octahedral (Octahedral), diamond (Diamond), and helical (Gyroid) structures were constructed with similar porosity and macropore diameter. CaP ceramics with the HCP structure exhibited higher compression strength (8.39 ± 1.82 MPa) and lower permeability (6.41 × 10−11 m2) compared to the Octahedral, Diamond, and Gyroid structures. In vitro cellular responses indicated that the macropore architecture strongly influenced the local growth rate of osteoblast-formed cell tissue; cells grew uniformly and formed circular rings in the HCP group. Furthermore, the HCP group promoted the expression of osteogenic genes and proteins more effectively than the other three groups. The outstanding osteoinductivity of the HCP group was confirmed in canine intramuscular implantation studies, where the new bone area reached up to 8.02 ± 1.94 % after a 10-week implantation. Additionally, the HCP group showed effective bone regeneration in repairing femoral condyle defects. Therefore, our findings suggest that 3D printed CaP bioceramics with an HCP structure promote osteoinductivity and can be considered as candidates for personalized precise treatment of bone defects in clinical applications. Statement of significance1. 3D printing BCP ceramics with high osteoinductivity were constructed through pore architecture optimization.2. BCP ceramics with HCP structure exhibited relatively higher mechanical strength and lower permeability than those with Octahedral, Diamond and Gyroid structures.3. BCP ceramics with HCP structure could promote the osteogenic differentiation of MC3T3-E1, and showed the superior in-vivo osteoinductivity and bone regeneration comparing with the other structures.

Indirect 3D printing CDHA scaffolds with hierarchical porous structure to promote osteoinductivity and bone regeneration

Hierarchical porous structure, which include macropores, minor pores, and micropores in scaffolds, are essential in the multiple biological functions of bone repair and regeneration. In this study, patient-customized calcium-deficient hydroxyapatite (CDHA) scaffolds with three-level hierarchical porous structure were fabricated by indirect 3D printing technology and particulate leaching method. The sacrificial template scaffolds were fabricated using a photo-curing 3D printer, which provided a prerequisite for the integral structure and interconnected macropores of CDHA scaffolds. Additionally, 20 wt% pore former was incorporated into the slurry to enhance the content of smaller pores within the CDHA-2 scaffolds, and then the CDHA-2 scaffolds were sintered to remove the sacrificial template scaffolds and pore former. The obtained CDHA-2 scaffolds exhibited interconnected macropores (300–400 μm), minor pores (∼10–100 μm), and micropores (< 10 μm) distributed throughout the scaffolds, which could promote bone tissue ingrowth, increase surface roughness, and enhance protein adsorption of scaffolds. In vitro studies identified that CDHA-2 scaffolds had nanocrystal grains, high specific surface area, and outstanding protein adsorption capacity, which could provide a microenvironment for cell adhesion, spreading, and proliferation. In addition, the murine intramuscular implantation experiment suggested that CDHA-2 scaffolds exhibited excellent osteoinductivity and were superior to traditional BCP ceramics under conditions without the addition of live cells and exogenous growth factors. The rabbit calvarial defect repair results indicated that CDHA-2 scaffolds could enhance in situ bone regeneration. In conclusion, these findings demonstrated that the hierarchical porous structure of CDHA scaffolds was a pivotal factor in modulating osteoinductivity and bone regeneration, and CDHA-2 scaffolds were potential candidates for bone regeneration.

Key informant views on potential acceptability and feasibility of long-acting antiretroviral treatment for HIV in Kenya

BackgroundIn 2020, 14% of diagnosed persons living with HIV (PLWH) in Kenya were not taking antiretroviral therapy (ART), and 19% of those on ART had unsuppressed viral loads. Long-acting antiretroviral therapy (LA-ART) may increase viral suppression by promoting ART uptake and adherence. We conducted key informant (KI) interviews with HIV experts in Kenya to identify product and delivery attributes related to the acceptability and feasibility of providing LA-ART to PLWH in Kenya.MethodsInterviews were conducted via Zoom on potential LA-ART options including intra-muscular (IM) injections, subcutaneous (SC) injections, implants, and LA oral pills. KI were asked to discuss the products they were most and least excited about, as well as barriers and facilitators to LA-ART roll-out. In addition, they were asked about potential delivery locations for LA-ART products such as homes, pharmacies, and clinics. Interviews were recorded and transcribed, and data were analyzed using a combination of inductive and deductive coding.ResultsTwelve KI (5 women, 7 men) participated between December 2021 and February 2022. Overall, participants reported that LA-ART would be acceptable and preferable to PLWH because of fatigue with daily oral pills. They viewed IM injections and LA oral pills as the most exciting options to ease pill burden and improve adherence. KI felt that populations who could benefit most were adolescents in boarding schools and stigmatized populations such as sex workers. SC injections and implants were less favored, as they would require new training initiatives for patients or healthcare workers on administration. In addition, SC injections would require refrigeration and needle disposal after use. Some KI thought patients, especially men, might worry that IM injections and implants would impact fertility, given their role in family planning. Pharmacies were perceived by most KI as suboptimal delivery locations; however, given ongoing work in Kenya to include pharmacies in antiretroviral delivery, they recommended asking patients their views.ConclusionThere is interest and support for LA-ART in Kenya, especially IM injections and LA oral pills. Identifying patient preferences for modes and delivery locations and addressing misconceptions about specific products as they become available will be important before wide-scale implementation.

Non-pyrogenicity and biocompatibility of parylene-coated magnetic bead implants.

Clinical grade magnetic bead implants have important applications in interfacing with the human body, providing contactless mechanical attachment or wireless communication through human tissue. We recently developed a new strategy, magnetomicrometry, that uses magnetic bead implants as passive communication devices to wirelessly sense muscle tissue lengths. We manufactured clinical-grade magnetic bead implants and verified their biocompatibility via intramuscular implantation, cytotoxicity, sensitization, and intracutaneous irritation testing. In this work, we test the pyrogenicity of the magnetic bead implants via a lagomorph model, and we test the biocompatibility of the magnetic bead implants via a full chemical characterization and toxicological risk assessment. Further, we test the cleaning, sterilization, and dry time of the devices that are used to deploy these magnetic bead implants. We find that the magnetic bead implants are non-pyrogenic and biocompatible, with the insertion device determined to be safe to clean, sterilize, and dry in a healthcare setting. These results provide confidence for the safe use of these magnetic bead implants in humans.

The effects of heat and hydrogen peroxide treatment on the osteoinductivity of demineralized cortical bone: a potential method for preparing tendon/ligament repair scaffolds.

Recent studies have indicated that demineralized cortical bone (DCB) may be used to repair tendons and ligaments, such as the patellar tendon and anterior cruciate ligament (ACL). Hydrogen peroxide (H2O2) has been shown to reduce the osteoinductivity of DCB, and heat treatment may also decrease the osteoinductivity of DCB. The purpose of this study was (i) to determine whether heat treatment reduces the osteoinductivity of DCB and (ii) to compare the effectiveness of heat treatment and H2O2 treatment on BMP-2 inactivation. DCB was prepared by immersion in 0.6 N hydrochloric acid, and DCB-H and DCB-HO were prepared by heat treatment (70°C for 8 h) and H2O2 treatment (3% H2O2 for 8 h), respectively. The surface topographies, elemental distributions and histological structures of the scaffolds were observed by scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FT-IR) and histological staining. The viability and osteogenic differentiation of TDSCs cultured on the scaffolds were evaluated via live/dead cell staining and Cell Counting Kit-8 (CCK-8) testing, real-time polymerase chain reaction (RT-PCR) and western bolt (WB) analysis, alkaline phosphatase activity (ALP) and alizarin red S (ARS) staining. The intramuscular implantation of the scaffolds in rats was also used to evaluate the effect of heat treatment and H2O2 treatment on the osteoinductivity of DCB. Our results demonstrated that both treatments removed BMP-2 and osteocalcin (OCN) within the DCB and that DCB-H and DCB-HO had good cytocompatibility and reduced the osteogenic differentiation of TDSCs. Moreover, the in vivo results indicated that the DCB-H and DCB-HO groups had smaller areas of osteoid formation than did the DCB group, and the DCB-HO group had the smallest area among the three groups. Our study demonstrated that heat treatment could reduce the osteoinductivity of DCB, and that H2O2 treatment was more effective than heat treatment.

PI3K/AKT/mTOR signaling regulates BCP ceramic-induced osteogenesis.

An increasing number of studies demonstrate that biphasic calcium phosphate (BCP) ceramics can induce bone regeneration. However, the underlying molecular mechanisms involved are still poorly understood. This work was proposed to investigate how PI3K/AKT/mTOR signaling influenced the osteogenesis mediated by BCP ceramics. The results showed that incubation with BCP ceramics promoted the proliferation of murine bone marrow-derived mesenchymal stem cells (BMSCs) in a time-dependent manner. The resulting cell proliferation was then suppressed by the selective inhibition of either PI3K, AKT, or mTOR signaling activation. Next, we confirmed that BCP ceramics up-regulated the phosphorylation levels of AKT and mTOR in BMSCs, suggesting the ability of BCP ceramics to drive the activation of PI3K/AKT/mTOR signaling in BMSCs. Furthermore, the blockade of PI3K/AKT/mTOR signaling prevented BCP ceramics-induced osteogenic differentiation and pro-angiogenesis of BMSCs by down-regulating the expression of genes encoding OPN, RUNX2 or VEGF. Moreover, the PI3K/AKT/mTOR signaling blockade suppressed stem cell infiltration and new bone formation in the implants following intra-muscular implantation of BCP ceramics in mice. Therefore, our results suggested that PI3K/AKT/mTOR signaling played a critical regulatory role in BCP ceramic-induced osteogenesis.

Hybrid implants based on calcium-magnesium silicate ceramic diopside as a carrier of recombinant BMP-2 and demineralized bone matrix as a scaffold: ectopic osteogenesis in intramuscular implantation in mice

High efficiency of hybrid implants based on calcium-magnesium silicate ceramic diopside as a carrier of recombinant BMP-2 and xenogenic demineralized bone matrix (DBM) as a scaffold for bone tissue regeneration was demonstrated previously on the model of critical size cranial defects in mice. In order to investigate the possibility of using these implants for growing autologous bone tissue using the in vivo bioreactor principle in the patient’s own body, the effectiveness of the ectopic osteogenesis induced by them in intramuscular implantation in mice was studied. At 7 µg dose of BMP-2 per implant, after 1 week we observed a dense agglomeration of cells, probably skeletal muscle satellite precursor cells, with areas of intense chondrogenesis, the initial stage of indirect osteogenesis, around the implants. After 12 weeks, a dense bone capsule of trabecular structure was formed, covered with periosteum, with mature bone marrow located in the spaces between the trabeculae. The capsule volume was about 8-10 times the volume of the original implant. There were practically no signs of inflammation and foreign body reaction. Microcomputer tomography data showed a significant increase of the relative bone volume, number of trabeculae and bone tissue density in the group with BMP-2 in comparison with the group without BMP-2. Considering that DBM can be obtained in practically unlimited quantities, of the required size and shape, and the BMP-2 used, which is obtained by synthesis in E. coli cells, is relatively inexpensive, further development of the in vivo bioreactor model based on hybrid implants from BMP-2, diopside and xenogenic DBM seems promising.

The genetic background determines material-induced bone formation through the macrophage-osteoclast axis

Osteoinductive materials are characterized by their ability to induce bone formation in ectopic sites. Thus, osteoinductive materials hold promising potential for repairing bone defects. However, the mechanism of material-induced bone formation remains unknown, which limits the design of highly potent osteoinductive materials. Here, we demonstrated a genetic background link among macrophage polarization, osteoclastogenesis and material-induced bone formation. The intramuscular implantation of an osteoinductive material in FVB/NCrl (FVB) mice resulted in more M2 macrophages at week 1, more osteoclasts at week 2 and increased bone formation after week 4 compared with the results obtained in C57BL/6JOlaHsd (C57) mice. Similarly, in vitro, with a greater potential to form M2 macrophages, monocytes derived from FVB mice formed more osteoclasts than those derived from C57 mice. A transcriptomic analysis identified Csf1, Cxcr4 and Tgfbr2 as the main genes controlling macrophage-osteoclast coupling, which were further confirmed by related inhibitors. With such coupling, macrophage polarization and osteoclast formation of monocytes in vitro successfully predicted in vivo bone formation in four other mouse strains. Considering material-induced bone formation as an example of acquired heterotopic bone formation, the current findings shed a light on precision medicine for both bone regeneration and the treatment of pathological heterotopic bone formation.

A Novel Two-Step Ultrasound-Guided Technique for Anesthesia and Postsurgery Analgesia in Gluteal Augmentation Surgery.

In recent years, there has been an increase in demand for gluteal augmentation and reshaping surgeries with intramuscular implants which are becoming increasingly popular. Until now, this surgery was mainly performed under general anesthesia, but recently locoregional anesthesia techniques, such as tumescent local anesthesia, are being applied more and more. Today, the use of ultrasound for locoregional anesthesia and analgesia allows us to perform techniques that are increasingly precise and burdened with lower risks. In this report, we present a novel two-step ultrasound-guided technique combining a botox injection in the gluteus maximus four weeks before surgery and tumescent anesthesia with a cluneal nerveblock.Furthermore, the combination of anesthetic and analgesic techniques can guarantee a better result both in terms of surgical execution of the interventionand in reducing pain and improving patient comfort in the postoperative period.

Hybrid Implants Based on Calcium-Magnesium Silicate Ceramic Diopside as a Carrier of Recombinant BMP-2 and Demineralized Bone Matrix as a Scaffold: Ectopic Osteogenesis in Intramuscular Implantation in Mice

High efficiency of hybrid implants based on calcium-magnesium silicate ceramic, diopside, as a carrier of recombinant BMP-2 and xenogenic demineralized bone matrix (DBM) as a scaffold for bone tissue regeneration was demonstrated previously using the model of critical size cranial defects in mice. In order to investigate the possibility of using these implants for growing autologous bone tissue using in vivo bioreactor principle in the patient’s own body, effectiveness of ectopic osteogenesis induced by them in intramuscular implantation in mice was studied. At the dose of 7 μg of BMP-2 per implant, dense agglomeration of cells, probably skeletal muscle satellite precursor cells, was observed one week after implantation with areas of intense chondrogenesis, initial stage of indirect osteogenesis, around the implants. After 12 weeks, a dense bone capsule of trabecular structure was formed covered with periosteum and mature bone marrow located in the spaces between the trabeculae. The capsule volume was about 8-10 times the volume of the original implant. There were practically no signs of inflammation and foreign body reaction. Microcomputed tomography data showed significant increase of the relative bone volume, number of trabeculae, and bone tissue density in the group of mice with BMP-2-containing implant in comparison with the group without BMP-2. Considering that DBM can be obtained in practically unlimited quantities with required size and shape, and that BMP-2 is obtained by synthesis in E. coli cells and is relatively inexpensive, further development of the in vivo bioreactor model based on the hybrid implants constructed from BMP-2, diopside, and xenogenic DBM seems promising.

Pmma body injection and hypercalcemia correlation: a cross-sectional observational study on the comprehensive analysis of variables

Introduction: Hypercalcemia related to aesthetic intramuscular implants exclusively made of PMMA (polymethylmethacrylate) is a very rare condition. Late complications of these implants are related to poorly understood inflammatory triggers, which may reactivate stabilized implants, increase calcitriol production, and lead to hypercalcemia. Methods: Records of patients seen between 2022 and 2023 with a PMMA microsphere body implant and a subsequent diagnosis of hypercalcemia were searched. Medical history, medication use, data on PMMA implantation, and laboratory tests related to calcium metabolism were described and subjected to variable correlation analysis. The instituted treatments were described. Results: A total of 17 female patients with exclusive body PMMA implants confirmed by ultrasonography or MRI (Magnetic Resonance Image) developed hypercalcemia between 2017 and 2022. Prior renal disease, COVID-19 infection, vitamin D use, and anabolic steroid use correlated with laboratory changes in hypercalcemia. Larger PMMA implant volumes led to parathyroid hormone suppression. Conclusion: Patients with PMMA body implants may rarely present with hypercalcemia when variables such as renal disease before implantation, high doses of vitamin D or anabolic steroids are present. Based on the data presented and the available scientific literature, recommendations for patient investigation, implant volume limit, and patient follow-up after the procedure were made.

Poster 105: Rotator cuff healing using demineralised bone fibers in ovine infraspinatus tendon models

Objectives: Rotator cuff tears are a common cause of shoulder pain and functional deficits. Although surgical repair is often able to restore function and resolve pain, high rates of postoperative structural failures have been reported. To improve the outcomes and reduce re-tear rates biologic tissue scaffolds have been used to augment and reinforce the primary tendon repair by providing mechanical support of the initial repair and ideally improving the rate and/or quality of biological healing at the tendon-bone interface. Demineralised bone fibers (DBF) are bone collagen and can be formed into a patch providing a scaffold and inductive bone morphogenetic proteins known to facilitate healing. This study evaluated the in vivo response of human DBF Patches (hDBF-P) as an onlay over the infraspinatus tendon (IST) and at the tendon-bone interface in an ovine model. Methods: The hDBF-P was characterised using stereozoom (Figure 1A) and environment electron microscopy (Figure 1B). The osteoinductivity (OI) was evaluated in the standard nude rat intramuscular implantation (IM) model at 28 days using mCT and paraffin histology. A bilateral model of an onlay or an enthesis repair was performed in 12 adult wethers following ethical approval. Four groups were evaluated at 6 and 12 weeks (n=3 per time point): Onlay control and hDBF treated; Enthesis repair control and hDBF treated. The onlay model simulated a tendon injury by 4 longitudinal slits along the IST and lateral drill holes in the proximal humerus in the onlay control group [1]. In the onlay treated group the hDBF-P was hydrated in saline and secured to the IST with interrupted 2-0 non-resorbable sutures (Figure 2A). On contralateral side, the central third of the IST was sharply dissected, the footprint burred, and 2 medial row anchors inserted. The sutures were passed through the IST in the enthesis repair control group. In the enthesis repair treatment group, the sutures were passed through the hDBF-P which was placed at the tendon bone interface followed by sutures through the IST. Two lateral push lock anchors were used for a suture bridge (Figure 2B) in the control and treated groups. Endpoints included clinical recovery, macroscopic dissection, 3T MRI, radiographs, mCT and paraffin histology processing, H&amp;E staining, and light microscopic evaluations to assess local cell and tissue responses and healing. Results: OI was confirmed on IM implantation in nude rats. The sheep recovered uneventfully with no adverse events macroscopically or radiographically. MRI in the onlay control group revealed little response and no increase in tendon thickness which was confirmed histologically. In contrast the onlay hDBF-P treated demonstrated increased tendon thickness and hDBF resorption with time. Histology showed some initial inflammatory cells that decrease with time in the onlay group. MRI in the hDBF-P treated enthesis group supported increased activity at the bone interface at 6 and 12 weeks compared to controls. Histology at the interface with hDBF-P showed evidence of new bone formation at 6 weeks that matured with time support enthesis reformation. Conclusions: hDBF-P is a collagen graft composed of demineralized bone fibers that have osteoconductive and osteoinductive potential. When formed into a patch, this implant provides a scaffold that can participate with the local biology. hDBF-P used as on-lay in a tendon injury model increased tendon thickness. hDBF-P when used at the interface between the tendon and bone facilitated enthesis reformation through its conductive and inductive potential as well as scaffold architecture. DBF patches represent an exciting allograft implant with the potential to address the clinical problems in rotator cuff surgery. Literature cited Van Kampen, C., et al. (2013). Tissue-engineered augmentation of a rotator cuff tendon using a reconstituted collagen scaffold: a histological evaluation in sheep. Muscles, ligaments and tendons journal, 3(3), p.229.

Becker's Nevus Syndrome - gluteal muscle asymmetry treatment with polymethylmethacrylate implant - a case report

Becker's nevus syndrome presents with a hyperchromic spot often trunk located, associated with hypertrichosis and maxillofacial or skeletal and muscle alterations. This study reports the case of a male patient who had Becker's nevus syndrome in the left gluteal region, who underwent intramuscular PMMA microspheres implantation for muscle volume symmetrization

Application of Exogenous GnRH in Food Animal Production.

Over several decades, exogenous GnRH and agonists have been employed for controlling reproductive cascades in animals, and treating some reproductive morbidities. The administration of GnRH is used in animals to counter ovarian dysfunction, induce ovulation, and to increase conception and pregnancy rates. GnRH and its agonists are used in the treatment of cystic ovarian degeneration and repeat breeder syndrome. The development of protocols for GnRH administration by intramuscular injection, intramuscular or subcutaneous implants, and intravaginal deposition has empowered their clinical use worldwide. Currently, exogenous GnRH products are a central part of several pre- and post-breeding programs for the enhancement of fertility, including the control of estrous cycles and timing of ovulation, development of fixed-time artificial insemination protocols, improved embryo survival, and the treatment of reproductive morbidity. The aim of the present review is to summarize the application of exogenous GnRH agonists in food animal production.

One-step co-doping of ZnO and Zn2+ in osteoinductive calcium phosphate ceramics with synergistic antibacterial activity for regenerative repair of infected bone defect

How to endow bone grafts with long-term antibacterial activity and good bone regenerative ability to achieve the regenerative repair of infected bone defects has been the focus of the clinical treatment of osteomyelitis. The present study introduced a novel one-step route to realizing the co-doping of zinc oxide (ZnO) and zinc ion (Zn2+) in biphasic calcium phosphate (BCP) ceramics to utilize their synergistic antibacterial. Compared with the conventional BCP ceramics (BCP-Ca), the ZnO/Zn2+ co-doping ones (BCP-Zn) possessed strong antibacterial ability on E. coli and S. aureus as well as stimulated the osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) effectively. The synergistic antibacterial mechanism of ZnO and Zn2+ was also investigated. BCP-Zn showed excellent osteoinductivity and angiogenesis at three months postoperatively in the canine intramuscular implantation model. Moreover, BCP-Zn exhibited excellent anti-infective ability and bone regenerative repair compared to BCP-Ca and control groups in the infected bone defect model of rat femur. Collectively, these findings suggest that the simultaneous introduction of ZnO/Zn2+ could have immense potential to expand the application of osteoinductive BCP ceramics in the regenerative repair of infected bone defects.

Calcium-to-phosphorus releasing ratio affects osteoinductivity and osteoconductivity of calcium phosphate bioceramics in bone tissue engineering

Calcium phosphate (Ca-P) bioceramics, including hydroxyapatite (HA), biphasic calcium phosphate (BCP), and beta-tricalcium phosphate (β-TCP), have been widely used in bone reconstruction. Many studies have focused on the osteoconductivity or osteoinductivity of Ca-P bioceramics, but the association between osteoconductivity and osteoinductivity is not well understood. In our study, the osteoconductivity of HA, BCP, and β-TCP was investigated based on the osteoblastic differentiation in vitro and in situ as well as calvarial defect repair in vivo, and osteoinductivity was evaluated by using pluripotent mesenchymal stem cells (MSCs) in vitro and heterotopic ossification in muscles in vivo. Our results showed that the cell viability, alkaline phosphatase activity, and expression of osteogenesis-related genes, including osteocalcin (Ocn), bone sialoprotein (Bsp), alpha-1 type I collagen (Col1a1), and runt-related transcription factor 2 (Runx2), of osteoblasts each ranked as BCP > β-TCP > HA, but the alkaline phosphatase activity and expression of osteogenic differentiation genes of MSCs each ranked as β-TCP > BCP > HA. Calvarial defect implantation of Ca-P bioceramics ranked as BCP > β-TCP ≥ HA, but intramuscular implantation ranked as β-TCP ≥ BCP > HA in vivo. Further investigation indicated that osteoconductivity and osteoinductivity are affected by the Ca/P ratio surrounding the Ca-P bioceramics. Thus, manipulating the appropriate calcium-to-phosphorus releasing ratio is a critical factor for determining the osteoinductivity of Ca-P bioceramics in bone tissue engineering.

3D-Printed Porous Tantalum Scaffold Improves Muscle Attachment via Integrin-β1-Activated AKT/MAPK Signaling Pathway.

3D-printed porous titanium (Ti) alloy scaffolds have been reported for facilitating muscle attachment in our previous study. However, the anti-avulsion ability needs to be improved. In this study, we used 3D-printed porous tantalum (Ta) scaffolds to improve muscle attachment. The differences in chemical and physical characteristics and muscle adhesion between the two scaffolds were tested and compared in the gene and protein level both in vitro and in vivo. The possible molecular mechanism was analyzed and further proved. The results showed that compared with the porous Ti alloy, porous Ta had better cell proliferation, differentiation, migration, and adhesion via the integrin-β1 (Itgb1)-activated AKT/MAPK signaling pathway in L6 rat myoblasts. When artificially down-regulated the expression of Itgb1, cell adhesion and myogenesis differentiation were affected and the phosphorylation of the AKT/MAPK signaling pathway was suppressed. In rat intramuscular implantation, porous Ta had a significantly higher muscle ingrowth rate (85.63% ± 4.97 vs 65.98% ± 4.52, p < 0.01) and larger avulsion force (0.972 vs 0.823 N/mm2, p < 0.05) than the porous Ti alloy. These findings demonstrate that the 3D-printed porous Ta scaffold is beneficial for further clinical application of muscle attachment.