Macrophage phenotype is determinant for fibrosis development in keloid disease.

Keloid refers to a fibroproliferative disorder characterized by an accumulation of extracellular matrix (ECM) components at the dermis level, overgrowth beyond initial wound, and formation of tumor-like nodule areas. Treating keloid is still an unmet clinical need and the lack of an efficient therapy is clearly related to limited knowledge about keloid etiology, despite the growing interest of the scientific community in this pathology. In past decades, keloids were often studied in vitro through the sole prism of fibroblasts considered as the major effector of ECM deposition. Nevertheless, development of keloids results from cross-interactions of keloid fibroblasts (KFs) and their surrounding microenvironment, including immune cells such as macrophages. Our study aimed to evaluate the effect of M1 and M2 monocyte-derived macrophages on KFs in vitro. We focused on the effects of the macrophage secretome on fibrosis-related criteria in KFs, including proliferation, migration, differentiation, and ECM synthesis. First, we demonstrated that M2-like macrophages enhanced the fibrogenic profile of KFs in culture. Then, we surprisingly founded that M1-like macrophages can have an anti-fibrogenic effect on KFs, even in a pro-fibrotic environment. These results demonstrate, for the first time, that M1 and M2 macrophage subsets differentially impact the fibrotic fate of KFs in vitro, and suggest that restoring the M1/M2 balance to favor M1 in keloids could be an efficient therapeutic lever to prevent or treat keloid fibrosis.

Is Spheroid a Relevant Model to Address Fibrogenesis in Keloid Research?

Keloid refers to a fibro-proliferative disorder characterized by an accumulation of extracellular matrix at the dermis level, overgrowing beyond the initial wound and forming tumor-like nodule areas. The absence of treatment for keloid is clearly related to limited knowledge about keloid etiology. In vitro, keloids were classically studied through fibroblasts monolayer culture, far from keloid in vivo complexity. Today, cell aggregates cultured as 3D spheroid have gained in popularity as new tools to mimic tissue in vitro. However, no previously published works on spheroids have specifically focused on keloids yet. Thus, we hypothesized that spheroids made of keloid fibroblasts (KFs) could be used to model fibrogenesis in vitro. Our objective was to qualify spheroids made from KFs and cultured in a basal or pro-fibrotic environment (+TGF-ß1). As major parameters for fibrogenesis assessment, we evaluated apoptosis, myofibroblast differentiation and response to TGF-ß1, extracellular matrix (ECM) synthesis, and ECM-related genes regulation in KFs spheroids. We surprisingly observed that fibrogenic features of KFs are strongly downregulated when cells are cultured in 3D. In conclusion, we believe that spheroid is not the most appropriate model to address fibrogenesis in keloid, but it constitutes an efficient model to study the deactivation of fibrotic cells.

Reporting Criteria for Clinical Trials on Medication-Related Osteonecrosis of the Jaw (MRONJ): A Review and Recommendations.

Medication-related osteonecrosis of the jaw (MRONJ) is a complication caused by anti-resorptive agents and anti-angiogenesis drugs. Since we wanted to write a protocol for a randomized clinical trial (RCT), we reviewed the literature for the essential information needed to estimate the size of the active patient population and measure the effects of therapeutics. At the same time, we designed a questionnaire intended for clinicians to collect detailed information about their practices. Twelve essential criteria and seven additional items were identified and compiled from 43 selected articles. Some of these criteria were incorporated in the questionnaire coupled with data on clinical practices. Our review found extensive missing data and a lack of consensus. For example, the success rate often combined MRONJ stages, diseases, and drug treatments. The occurrence date and evaluation methods were not harmonized or quantitative enough. The primary and secondary endpoints, failure definition, and date coupled to bone measurements were not well established. This information is critical for writing a RCT protocol. With this review article, we aim to encourage authors to contribute all their findings in the field to bridge the current knowledge gap and provide a stronger database for the coming years.

Human amniotic membrane application in oral surgery-An ex vivo pilot study.

Objectives: The purpose of this pilot porcine study was to explore and illustrate the surgical application of human amniotic membrane (hAM) in an ex vivo model of medication-related osteonecrosis of the jaw (MRONJ). Material and methods: Five oral and maxillofacial surgeons participated to this study. MRONJ was simulated on porcine mandible specimens. hAM was applied using four different techniques: implantation with complete coverage, implantation with partial coverage, apposition and covering graft material. At the same time, the surgeons evaluated how well the hAM handled and its physical properties during the surgery. Results: Surgeons found that hAM had suitable mechanical properties, as it was easy to detach from the support, handle, bind to the defect and bury. hAM was also found to be strong and stable. The "implantation with complete coverage" and "implantation with partial coverage" techniques were the preferred choices for the MRONJ indication. Conclusion: This study shows that hAM is a graft material with suitable properties for oral surgery. It is preferable to use it buried under the gingiva with sutures above it, which increases its stability. This technical note aims to educate surgeons and provide them with details about the handling of hAM in oral surgery. Clinical relevance: Two surgical techniques for hAM application in MRONJ were identified and illustrated. hAM handling and physical properties during surgery were reported.

Does Penicillin Allergy Increase the Risk of Surgical Site Infection after Orthognathic Surgery? A Multivariate Analysis.

This study aimed to demonstrate an association between the occurrence of surgical site infection (SSI) after orthognathic surgery and penicillin allergy and to assess whether other factors could be associated with the occurrence of SSI. A 10-year monocentric retrospective study was conducted to identify possible risk factors for SSI in orthognathic surgery. Bivariate analyses were performed using Fisher, Student, or Wilcoxon tests and multivariate analyses using logistic regression. Two hundred and sixty-six patients were included, and 3.5% had SSI. Bivariate analyses revealed a significant association between SSI and age at surgery (p = 0.01), penicillin allergy (p = 0.02), and postoperative antibiotic therapy by Clindamycin (Dalacine®) (p = 0.02). Multivariate analyses confirmed the association between the occurrence of SSI and treatment with Clindamycin (Dalacine®) or Clindamycin (Dalacine®) and Metronidazole (Flagyl®) postoperatively (p = 0.04). Antibiotic therapy with Clindamycin (Dalacine®) seems to be associated with a higher rate of SSI, and the mandible was the only site affected by SSI.

Tips and Tricks and Clinical Outcome of Cryopreserved Human Amniotic Membrane Application for the Management of Medication-Related Osteonecrosis of the Jaw (MRONJ): A Pilot Study.

Medication-related osteonecrosis of the jaw (MRONJ) is a complication of certain pharmacological treatments such as bisphosphonates, denosumab, and angiogenesis inhibitors. There are currently no guidelines on its management, particularly in advanced stages. The human amniotic membrane (hAM) has low immunogenicity and exerts anti-inflammatory, antifibrotic, antimicrobial, antiviral, and analgesic effects. It is a source of stem cells and growth factors promoting tissue regeneration. hAM acts as an anatomical barrier with suitable mechanical properties (permeability, stability, elasticity, flexibility, and resorbability) to prevent the proliferation of fibrous tissue and promote early neovascularization at the surgical site. In oral surgery, hAM stimulates healing and facilitates the proliferation and differentiation of epithelial cells in the oral mucosa and therefore its regeneration. We proposed using cryopreserved hAM to eight patients suffering from cancer (11 lesions) with stage 2-3 MRONJ on a compassionate use basis. A collagen sponge was added in some cases to facilitate hAM grafting. One or three hAMs were applied and one patient had a reapplication. Three patients had complete closure of the surgical site with proper epithelialization at 2 weeks, and two of them maintained it until the last follow-up. At 1 week after surgery, three patients had partial wound dehiscence with partial healing 3 months later and two patients had complete wound dehiscence. hAM reapplication led to complete healing. All patients remained asymptomatic with excellent immediate significant pain relief, no infections, and a truly positive impact on the patients' quality of life. No adverse events occurred. At 6 months of follow-up, 80% of lesions had complete or partial wound healing (30 and 50%, respectively), while 62.5% of patients were in stage 3. Radiological evaluations found that 85.7% of patients had stable bone lesions (n = 5) or new bone formation (n = 1). One patient had a worsening MRONJ but remained asymptomatic. One patient did not attend his follow-up radiological examination. For the first time, this prospective pilot study extensively illustrates both the handling and surgical application of hAM in MRONJ, its possible association with a collagen sponge scaffold, its outcome at the site, the application of multiple hAM patches at the same time, and its reapplication.

Surgical Application of Human Amniotic Membrane and Amnion-Chorion Membrane in the Oral Cavity and Efficacy Evaluation: Corollary With Ophthalmological and Wound Healing Experiences.

Due to its intrinsic properties, there has been growing interest in human amniotic membrane (hAM) in recent years particularly for the treatment of ocular surface disorders and for wound healing. Herein, we investigate the potential use of hAM and amnion-chorion membrane (ACM) in oral surgery. Based on our analysis of the literature, it appears that their applications are very poorly defined. There are two options: implantation or use as a cover material graft. The oral cavity is submitted to various mechanical and biological stimulations that impair membrane stability and maintenance. Thus, some devices have been combined with the graft to secure its positioning and protect it in this location. This current opinion paper addresses in detail suitable procedures for hAM and ACM utilization in soft and hard tissue reconstruction in the oral cavity. We address their implantation and/or use as a covering, storage format, application side, size and number, multilayer use or folding, suture or use of additional protective covers, re-application and resorption/fate. We gathered evidence on pre- and post-surgical care and evaluation tools. Finally, we integrated ophthalmological and wound healing practices into the collected information. This review aims to help practitioners and researchers better understand the application of hAM and ACM in the oral cavity, a place less easily accessible than ocular or cutaneous surfaces. Additionally, it could be a useful reference in the generation of new ideas for the development of innovative protective covering, suturing or handling devices in this specific indication. Finally, this overview could be considered as a position paper to guide investigators to fulfill all the identified criteria in the future.

Halofuginone regulates keloid fibroblast fibrotic response to TGF-ß induction.

Keloids are characterized by increased deposition of fibrous tissue in the skin and subcutaneous tissue following an abnormal wound healing process. Although keloid etiology is yet to be fully understood, fibroblasts are known to be key players in its development. Here we analyze the antifibrotic mechanisms of Halofuginone (HF), a drug reportedly able to inhibit the TGF-ß1-Smad3 pathway and to attenuate collagen synthesis, in an in-vitro keloid model using patient-derived Keloid Fibroblasts (KFs) isolated from fibrotic tissue collected during the "Scar Wars" clinical study (NCT NCT03312166). TGF-ß1 was used as a pro-fibrotic agent to stimulate fibroblasts response under HF treatment. The fibrotic related properties of KFs, including survival, migration, proliferation, myofibroblasts conversion, ECM synthesis and remodeling, were investigated in 2D and 3D cultures. HF at 50 nM concentration impaired KFs proliferation, and decreased TGF-ß1-induced expression of α-SMA and type I procollagen production. HF treatment also reduced KFs migration, prevented matrix contraction and increased the metallo-proteases/inhibitors (MMP/TIMP) ratio. Overall, HF elicits an anti-fibrotic contrasting the TGF-ß1 stimulation of KFs, thus supporting its therapeutic use for keloid prevention and management.

Impact of intraoperative cone beam computed tomography in the management of zygomatic fractures.

AIM: To evaluate the impact of intraoperative cone beam computed tomography (CBCT) assessment for the surgical management of zygomatic bone fracture (ZF). METHODS: A prospective study was conducted on patients operated on for unilateral ZF during a two-year period. Repositioning of the fractured zygoma was assessed intraoperatively by measuring the position of malar eminences in three dimensions on CBCT. A difference of more than 2 mm between both sides was considered significant and the reduction was judged inadequate. Surgical procedure was adapted to intraoperative imaging findings. RESULTS: The surgical procedures of 47 patients treated for ZF were analyzed. In 15% of the cases (7 patients), the intraoperative CBCT showed an inadequate reduction while the morphological results seemed correct. Those patients benefited from an immediate correction of the reduction. From these 7 patients, reduction was optimized further without the need of osteosynthesis in one case, additional osteosynthesis was performed in 4 cases and 2 patients required both reduction and osteosynthesis revision. CONCLUSION: Intraoperative CBCT control helps to achieve anatomic repositioning in case of ZF. It may reduce the risk of under-treatment and possible reoperation, and of over-treatment meaning systematic ORIF in all fractures.

Multimodal investigation of a keloid scar by combining mechanical tests in vivo with diverse imaging techniques.

Keloids are pathologic scars, defined as fibroproliferative diseases resulting from abnormal wound responses, which grow beyond the original wound margins. They develop on specific pro-keloid anatomic sites frequently characterized by high stress states. The initiation and growth mechanisms of keloid are not well-understood. This study relates multimodal investigation of a keloid by using mechanical tests in vivo and imaging techniques. A single case composed of a keloid, the healthy skin surrounding the keloid, and the contralateral healthy skin on the upper arms of a woman has been investigated in extension and suction by using non-invasive devices dedicated to in vivo skin measurement. The thickness and microstructure of these soft tissues have been observed by echography, tomography and confocal microscopy. Displacement fields have been obtained by using digital image correlation. Unlike healthy skin, keloid is not a well-defined multilayer structure: the frontier between epidermis and dermis disappears. The mechanical behavior of keloid is highly different from healthy skin one. The R-parameters have been deduced from suction curves. Physical parameters as tissue extensibility, initial and final tangent moduli have been identified from the stress-strain curves. The extensibility (respectively, initial rigidity) of keloid is highly lower (respectively, higher) than that of healthy skin. To compare the final rigidity of keloid versus healthy skin, further tests have to be performed with higher strain values.

Surgical guide for the remodelling of the orbito-naso-frontal bandeau in craniosynostosis surgery.

INTRODUCTION: Craniosynostoses affecting the forehead sutures can not only cause brain damage, but can also have an esthetic impact, because of the associated orbito-naso-frontal deformations. Reshaping the orbito-naso-frontal bandeau (ONFB) is difficult to appreciate perioperatively and should ideally be customized to each child. The aim of this study was to develop a template to guide the surgeon preoperatively towards an ideal customized remodelling of the ONFB. MATERIALS AND METHODS: A previous study conducted on computed tomography scans obtained from healthy children allowed us to conclude that the whole ONFB shape could be accurately described just by the distance measured between the fronto-zygomatic sutures (FZD), independently of age and gender. Our customizable template relies on this measurement. RESULTS: A re-usable template, built around three supports adjustable to a wide range of FZD, was designed using the CAD 3D Rhinoceros® software and machined in stainless steel 316L. The prototype was used for three children with good preliminary results. DISCUSSION AND CONCLUSION: The use of a customizable surgical template allows the surgeon to perform accurate and ideal perioperative remodeling of the ONFB in children suffering from craniosynostosis affecting the forehead sutures. Our prototype is currently the only one to be adjustable according to the FZD. The utility of this device will be assessed in a future prospective clinical study.

Clinical experience with osteosynthesis of subcondylar fractures of the mandible using TCP plates.

AIM: To evaluate the clinical and radiological results obtained with a new kind of osteosynthesis device (Modus TCP) plates, Medartis, Basel, Switzerland), especially designed for low subcondylar fracture (LSCF) and high subcondylar fracture (HSCF) of the mandible in association with the high submandibular approach (HSMA). METHOD: A prospective clinical and radiological study was carried out over a 41-month period. All adult patients suffering from a displaced LSCF or HSCF who consented to the surgical treatment were included in the study. All fractures were operated on using an HSMA and were stabilised using a 4- or 9-hole TCP((R)) plate. No intermaxillary fixation was used and a soft diet was started on the 1st postoperative day. Clinical examinations and control X-rays were carried out on the 10th postoperative day, in the 1st, the 3rd and, at least, in the 6th postoperative months. MATERIAL: Sixty-four patients (12 females, 52 males - mean age: 28.3) with a total of 75 fractures (54 LSCF, 21 HSCF) were included in the study. Mean postoperative follow-up was 14 months. RESULTS: In the 6th postoperative month, all fractures were consolidated in 79% of the cases in an anatomical position. A secondary displacement of the fracture occurred in 6.6% of the cases. Plate fracture was not observed. Dental occlusion remained unchanged in 94% of the patients. The mandibular movements (mouth opening, protrusion, lateral movements on the unfractured and fractured side) had normal mean values (49.5, 10.4, 12.8, 12.1mm, respectively). These movements were symmetrical in 95% of the patients. No patient complained about articular pain or noise. No facial nerve palsy was noticed, not even a transiently one. All patients but one found the scars acceptable. CONCLUSION: TCP plates, in association with HSMA, were found to be an efficient osteosynthesis device for stabilising subcondylar fractures.