Early Healing Phase Research Articles

Postoperative healing and complications based on anterior cruciate ligament reconstruction graft type.

Injury to the anterior cruciate ligament (ACL) is a devastating injury to athletes of all ages. The current gold standard treatment following complete rupture of the ACL is reconstruction of the torn ligament with autograft or allograft tendon. Commonly used tendon grafts include patellar tendon, hamstring tendon, and quadriceps tendon. Although ligaments and tendons have similar collagen and proteoglycan compositions, they maintain a unique composition and arrangement of cells to serve their unique biomechanical needs. Therefore, following ACL reconstruction (ACLR), the implanted tendon tissue undergoes a process of remodeling which is termed "ligamentization". The process of ligamentization is divided into three main phases, which include the early healing phase, the proliferative phase, and the maturation phase. Following the process of ligamentization, the graft tissue closely mimics the appearance of ligament tissue on an ultrastructural level. Successful outcome following ACLR is contingent upon adequate remodeling of the tissue as well as healing of the graft within the bone tunnels in the femur and tibia. Choice of graft has individual implications regarding their associated risk of complications, failure, and infection. The purpose of this review is to summarize the process of ligamentization and graft healing and to discuss how graft type influences the rate and types of complications, failures, and infections.

Bone healing, tissue effects and biomechanical fixation of ‘smooth’ ceramic-coated zirconia-based dental implants: An in vivo study in sheep

In this study, the effect of coating a zirconia-based ceramic oral implant with a material of the same composition to build a relatively smooth surface with three different porosity features was evaluated in vivo, at 4 and 8 weeks after implantation in sheep femoral condyles. The results showed that at 4 weeks, the three coated zirconia-based implants with smoother surface topographies behaved similarly and promoted faster bone healing compared to the results obtained in the same zirconia- or titanium-based implants, but with rougher sandblasted and acid-etched surfaces. In addition, higher pull-out strengths were estimated in the coated-ceramic sample compared to titanium sandblasted and etched one. The present work showed that zirconia coatings with smoother surfaces than those conventionally used in the market improved the early phase of bone healing, paving the way for shorter treatment times and improved patient outcomes.

Effects of the GingivalStat approach on early gingival margin remodeling following esthetic clinical crown lengthening: A case series.

The aim of this case series is to present the potential applications of the GingivalStat approach, that is, the use of temporary gingival stabilizers, to favor early gingival margin remodeling and prevent the occurrence of gingival rebound following esthetic clinical crown lengthening. Four patients requiring clinical crown lengthening were treated for esthetical and functional reasons. The surgical approach included: (a) gingival margin recontouring; (b) full-thickness flap elevation; (c) osteotomy (to achieve an adequate dimension between the alveolar bone crest and the CEJ) and osteoplasty (to reduce the bone thickness and improve the buccal bone anatomic profile, where indicated); (d) temporary gingival stabilizer placement using a block-out resin or a composite (the GingivalStat approach); and (e) flap repositioning, adaptation, and suture. One- to five-year follow-ups, reported in the different case scenarios, show evidence of clinically stable gingival margins around the treated teeth. Within the limits of this case series, it can be concluded that the GingivalStat approach appears as a further maneuver to cope with clinical crown lengthening procedures at esthetic sites. GingivalStat seems to favor gingival margin contour remodeling during the early phase of healing as well as prevent the occurrence of gingival rebound. GingivalStat approach may guide gingival margin remodeling and prevent gingival rebound after wound healing of sites submitted to esthetic clinical crown lengthening.

Review of Intraoral Vasculature and Implications on Incision Designs of Periodontal and Implant Surgeries.

Currently, the incision design for periodontal and implant surgeries is mainly based on the surgeon's personal preference. The primary aim of this study is to review the intricate periodontal microvascular system and to illustrate the potential impact of commonly applied flap designs on the integrity of this system. A complete literature electronic search resulted in 37 relevant articles. The maxillary, facial, and lingual arteries supply the microvasculature system, including the supraperiosteal, intraligamental, and intraosseous arterioles. These arterioles have their own territories yet are extensively interconnected. The impact of common papilla management techniques are discussed, including tunneling; papilla base, incision, and preservation; and flap-releasing incisions. Flap design can impact reperfusion and blood inflow in the early healing phase, which eventually influences wound closure probability, healing speed, surgical predictability, tissue volume change, and postoperative morbidity. Future studies on the three-dimensional distribution of microvasculature and clinical impact of various flap designs on tissue reperfusion can lead to evidence-based incision selection and improved wound-healing outcomes.

Suppressors of cGAS-STING are downregulated during fin-limb regeneration and aging in aquatic vertebrates.

During the early stages of limb and fin regeneration in aquatic vertebrates (i.e., fishes and amphibians), blastema undergo transcriptional rewiring of innate immune signaling pathways to promote immune cell recruitment. In mammals, a fundamental component of innate immune signaling is the cytosolic DNA sensing pathway, cGAS-STING. However, to what extent the cGAS-STING pathway influences regeneration in aquatic anamniotes is unknown. In jawed vertebrates, negative regulation of cGAS-STING activity is accomplished by suppressors of cytosolic DNA such as Trex1, Pml, and PML-like exon 9 (Plex9) exonucleases. Here, we examine the expression of these suppressors of cGAS-STING, as well as inflammatory genes and cGAS activity during caudal fin and limb regeneration using the spotted gar (Lepisosteus oculatus) and axolotl (Ambystoma mexicanum) model species, and during age-related senescence in zebrafish (Danio rerio). In the regenerative blastema of wounded gar and axolotl, we observe increased inflammatory gene expression, including interferon genes and interleukins 6 and 8. We also observed a decrease in axolotl Trex1 and gar pml expression during the early phases of wound healing which correlates with a dramatic increase in cGAS activity. In contrast, the plex9.1 gene does not change in expression during wound healing in gar. However, we observed decreased expression of plex9.1 in the senescing cardiac tissue of aged zebrafish, where 2'3'-cGAMP levels are elevated. Finally, we demonstrate a similar pattern of Trex1, pml, and plex9.1 gene regulation across species in response to exogenous 2'3'-cGAMP. Thus, during the early stages of limb-fin regeneration, Pml, Trex1, and Plex9.1 exonucleases are downregulated, presumably to allow an evolutionarily ancient cGAS-STING activity to promote inflammation and the recruitment of immune cells.

Distinct defects in early innate and late adaptive immune responses typify impaired fracture healing in diet-induced obesity.

Bone fractures, the most common musculoskeletal injuries, heal through three main phases: inflammatory, repair, and remodeling. Around 10% of fracture patients suffer from impaired healing that requires surgical intervention, a huge burden on the healthcare system. The rate of impaired healing increases with metabolic diseases such as obesity-associated hyperglycemia/type 2 diabetes (T2D), an increasing concern given the growing incidence of obesity/T2D. Immune cells play pivotal roles in fracture healing, and obesity/T2D is associated with defective immune-cell functions. However, there is a gap in knowledge regarding the stoichiometry of immune cells that populate the callus and how that population changes during different phases of healing. Here, we used complementary global and single-cell techniques to characterize the repertoire of immune cells in the fracture callus and to identify populations specifically enriched in the fracture callus relative to the unfractured bone or bone marrow. Our analyses identified two clear waves of immune-cell infiltration into the callus: the first wave occurs during the early inflammatory phase of fracture healing, while the second takes place during the late repair/early remodeling phase, which is consistent with previous publications. Comprehensive analysis of each wave revealed that innate immune cells were activated during the early inflammatory phase, but in later phases they returned to homeostatic numbers and activation levels. Of the innate immune cells, distinct subsets of activated dendritic cells were particularly enriched in the inflammatory healing hematoma. In contrast to innate cells, lymphocytes, including B and T cells, were enriched and activated in the callus primarily during the late repair phase. The Diet-Induced Obesity (DIO) mouse, an established model of obesity-associated hyperglycemia and insulin resistance, suffers from multiple healing defects. Our data demonstrate that DIO mice exhibit dysregulated innate immune responses during the inflammatory phase, and defects in all lymphocyte compartments during the late repair phase. Taken together, our data characterize, for the first time, immune populations that are enriched/activated in the callus during two distinct phases of fracture healing and identify defects in the healing-associated immune response in DIO mice, which will facilitate future development of immunomodulatory therapeutics for impaired fracture healing.

Dietary supplementation with a multi-strain yeast fraction enhances skin wound healing in zebrafish (Danio rerio)

Mechanical skin lesions are a persistent issue under aquaculture operations compromising fish health and robustness. Using an established zebrafish model, the study compared the potential skin wound healing benefit of dietary supplementation with a single-strain yeast fraction rich in β-glucans (β-glucan diet) or with a multi-strain yeast fraction (MsYF). Adult zebrafish were divided into four triplicated groups (12 tanks; 25 fish/tank) as negative control (unwounded-basal diet); control (wounded-basal diet), β-glucan (wounded-β-glucan diet), and MsYF (wounded-MsYF diet) and, following 7-week of nutritional preparation, inflicted with a full-thickness skin wound using a biopsy punch. Wound healing was assessed macroscopically by measuring wound surface area (18 fish individually tracked/group) until full healing within 4 weeks; as well as by histopathological diagnostic, transcriptional and immunoblotting analysis at the wound site at 1, 4 and 9 day post-wound (dpw) over the early healing phase (re-epithelization and inflammation). Following an initial expansion, wound closure was first measured at 4 dpw in the β-glucan and MsYF groups showing a positive and significantly higher daily wound closure rate compared to the control. Later at 16 dpw, cumulative and daily wound closure were significantly higher in the MsYF compared to the β-glucan and control groups. Enhanced gross wound healing was confirmed by histopathological diagnosis of mitigated inflammation, earlier re-epithelization and significantly enhanced granulation tissue synthesis in the MsYF group. Targeted transcriptomic and proteomic markers documented an early up-regulation of matrix metalloproteinases and a dampening or quicker resolution of inflammatory and cellular stress markers in the healing tissues of the MsYF compared to the control group while the β-glucan diet overall elicited intermediary results. Overall, this study demonstrates that dietary supplementation with a yeast-based functional compound can modulate fish immune response in wound healing and promote early wound closure indicating a role for skin health management under aquaculture operations.

Influence of surface nanotopography and wettability on early phases of peri-implant soft tissue healing: an in-vivo study in dogs

BackgroundIt is well established that nanotopography and wettability of implant surfaces contribute to osseointegration and long-term implant success. However, the effects of a hydrogenated surface with nanotubular and superhydrophilic properties on peri-implant soft tissue remain unclear. This study was designed to study the impact of a modified abutment surface on early soft tissue integration compared with a machined surface.MethodsThirty-six implants were placed at the bone level in the bilateral mandible of six beagles, followed by healing abutments belonging to the standard machined Ti-6Al-4V alloy abutments (TC4-M), anodized abutments with nanotubes (TC4-Nano), and hydrogenated abutments (TC4-H/Nano) groups, which were randomly screwed to the implants. After two and four weeks of wound healing, the animals were euthanized for histological evaluation.ResultsA superhydrophilic nanotubular surface developed on the hydrogenated abutment. Histological and histometric analyses revealed similar peri-implant soft tissue healing and dimensions for the three types of abutments at two and four weeks. Connective tissue (CT) length was longer around TC4-H/Nano abutments compared with standard abutments; however, the differences were not statistically significant. Moreover, collagen fibers in the TC4-H/Nano group extended and were attached perpendicularly to the superhydrophilic surface.ConclusionsOur results revealed that the soft tissue interface adjacent to the hydrogenated abutment is comparable to that of the machined abutment. A tendency of increased CT length and perpendicular collagen fibers was observed around the modified abutment. This study suggests that nanotubular/superhydrophilic surfaces could be a promising modification to enhance soft tissue sealing. However, comprehensive studies should be conducted to evaluate the peri-implant soft tissue around the modified abutment immunohistochemically, histopathologically, and clinically.

Electron Microscopy (EM) Analysis of Collagen Fibers in the Peri-Implant Soft Tissues around Two Different Abutments.

The design of the implant prosthesis-abutment complex appears crucial for shaping healthy and stable peri-implant soft tissues. The aim of the present animal study was to compare two implants with different healing abutment geometries: a concave design (TEST) and a straight one (CTRL). Transmission electron microscopy (TEM) was used to quantify the three-dimensional topography and morphological properties of collagen at nanoscale resolution. 2 swine were included in the experiment and 6 implants per animal were randomly placed in the left or right hemimandible in either the physiologically mature bone present between the lower canine and first premolar or in the mandibular premolar area, within tooth extraction sites. Each CTRL implant was positioned across from its respective TEST implant on the other side of the jaw. After 12 weeks of healing, 8 specimens (4 CTRL and 4 TEST) were retrieved and prepared for histological and TEM analysis. The results showed a significantly higher percentage of area covered by collagen bundles and average bundle size in TEST implants, as well as a significant decrease in the number of longitudinally oriented bundles with respect to CTRL implants, which is potentially due to the larger size of TEST bundles. These data suggest that a concave transmucosal abutment design serves as a scaffold, favoring the deposition and growth of a well-organized peri-implant collagen structure over the implant platform in the early healing phase, also promoting the convergence of collagen fibers toward the abutment collar.

Complications following alveolar ridge augmentation procedures.

Oral rehabilitation through implant supported dental restorations often requires a ridge augmentation procedure (RAP) prior to implant fixture placement since tooth extraction/loss results in alveolar ridge deficiencies. Although RAP-related surgical techniques and biomaterials have been in practice for several decades, outcomes are not always predictable. Post-surgical complications experienced during the early or late wound healing phases may jeopardize the targeted ideal ridge dimensions, required for implant fixture placement, and may have other consequences, such as negatively impacting the patient's quality of life. This review describes reported post-surgical complications following RAP under the following subtitles: complications by tissue type, complications in function and aesthetics, complications by healing time, complications by biomaterial type, and complications by surgical protocol modalities. Specifically, RAP performed by using particulate bone graft substitutes and related complications are explored. Modalities developed to prevent/manage these complications are also discussed.

Modified gelatin hydrogel nonwoven fabrics (Genocel) as a skin substitute in murine skin defects

From previous research, an emerging material composed of gelatin hydrogel nonwoven fabric (Genocel) has shown potential as a skin substitute, by improving neovascularization promotion in the early phase of wound healing. However, Genocel was inferior in terms of granulation formation compared to Pelnac. To solve this problem, we modified the manufacturing process of Genocel to reduce its water content, extend the degradation time (Genocel-L), and evaluate its healing process as a skin substitute. Genocel with a low water content (Genocel-L) was prepared and the difference in water content compared to that of the conventional Genocel was confirmed. Degradation tests were performed using collagenase and compared among Genocel-L, Genocel, and Pelnac sheets. In the invivo study, sheets of Genocel-L or Pelnac were applied to skin defects created on the backs of C57BL/6JJcl mice. On days 7, 14, and 21, the remaining wound area was evaluated and specimens were harvested for Hematoxylin and Eosin, Azan, anti-CD31, CD68, and CD163 staining to assess neoepithelialization, granulation tissue, capillary formation, and macrophage infiltration. Genocel-L had a lower water content than the conventional Genocel and a slower degradation than Genocel and Pelnac. In the invivo experiment, no significant differences were observed between Genocel-L and Pelnac in relation to the wound area, neoepithelium length, granulation formation, and the number of newly formed capillaries. The area of newly formed capillaries in the Pelnac group was significantly larger than that in the Genocel-L group on day 21 (p<0.05). Regarding macrophage infiltration, significantly more M2 macrophages were induced in the Pelnac group on days 14 and 21, and the M2 ratio was larger in the Pelnac group (p<0.05) during the entire process. Genocel-L has a lower water content and slower degradation rate than the conventional Genocel. Genocel-L had equivalent efficacy as a skin substitute to Pelnac, and can therefore be considered feasible for use as a skin substitute. However, a manufacturing method that can further modify Genocel-L is required to recover its early angiogenic potential.

Potential Delayed Healing in Wounds with Scabies Comorbidity: A Review about Immunological State and Oxidative Stress in Scabies Infestation

Scabies is a highly contagious skin disease caused by the human itch mite Sarcoptes scabiei var. hominid. Having scabies infestation coexisting in the same human body with a wound potentially lead to delayed wound healing. We constituted this literature review discussing wound healing in relation woth immunological state and oxidative stress in scabies infestation using information from literatures obtained from Google Scholar and PubMed. The immune response to scabies infection involves multiple layers of defense mechanisms, including the skin's defense mechanisms, T-cells, keratinocytes, and cytokines. Scabies mites can trigger upregulation of pro-inflammatory cytokines interleukins-1α (IL-1α) and interleukins-1β (IL-1β), which play a critical role in the early phases of wound healing by promoting the recruitment of immune cells and stimulating the production of growth factors and extracellular matrix proteins. Scabies mite infestation also triggers a sustained release of reactive oxygen species (ROS) from neutrophils which later becomes an oxidative stress leading to delayed healing and chronic inflammation. Topical treatment for scabies usually involves pesticide substances which, despite its therapeutic effect, have side effect of triggering ROS production. Comprehending the immunological state and oxidative stress in scabies infestation is essential in planning proper management for every wound with scabies comorbidity.

Diclofenac, a NSAID, delays fracture healing in aged mice

Nonsteroidal anti-inflammatory drugs (NSAIDs), such as diclofenac, belong to the most prescribed analgesic medication after traumatic injuries. However, there is accumulating evidence that NSAIDs impair fracture healing. Because bone regeneration in aged patients is subject to significant changes in cell differentiation and proliferation as well as a markedly altered pharmacological action of drugs, we herein analyzed the effects of diclofenac on bone healing in aged mice using a stable closed femoral facture model. Thirty-three mice (male n = 14, female n = 19) received a daily intraperitoneal injection of diclofenac (5 mg/kg body weight). Vehicle-treated mice (n = 29; male n = 13, female n = 16) served as controls. Fractured mice femora were analyzed by means of X-ray, biomechanics, micro computed tomography (μCT), histology and Western blotting. Biomechanical analyses revealed a significantly reduced bending stiffness in diclofenac-treated animals at 5 weeks after fracture when compared to vehicle-treated controls. Moreover, the callus tissue in diclofenac-treated aged animals exhibited a significantly reduced amount of bone tissue and higher amounts of fibrous tissue. Further histological analyses demonstrated less lamellar bone after diclofenac treatment, indicating a delay in callus remodeling. This was associated with a decreased number of osteoclasts and an increased expression of osteoprotegerin (OPG) during the early phase of fracture healing. These findings indicate that diclofenac delays fracture healing in aged mice by affecting osteogenic growth factor expression and bone formation as well as osteoclast activity and callus remodeling.

Spatiotemporal and microstructural characterization of heterotopic ossification in healing rat Achilles tendons.

Achilles tendon rupture is a common debilitating medical condition. The healing process is slow and can be affected by heterotopic ossification (HO), which occurs when pathologic bone-like tissue is deposited instead of the soft collagenous tendon tissue. Little is known about the temporal and spatial progression of HO during Achilles tendon healing. In this study we characterize HO deposition, microstructure, and location at different stages of healing in a rat model. We use phase contrast-enhanced synchrotron microtomography, a state-of-the-art technique that allows 3D imaging at high-resolution of soft biological tissues without invasive or time-consuming sample preparation. The results increase our understanding of HO deposition, from the early inflammatory phase of tendon healing, by showing that the deposition is initiated as early as one week after injury in the distal stump and mostly growing on preinjury HO deposits. Later, more deposits form first in the stumps and then all over the tendon callus, merging into large, calcified structures, which occupy up to 10% of the tendon volume. The HOs were characterized by a looser connective trabecular-like structure and a proteoglycan-rich matrix containing chondrocyte-like cells with lacunae. The study shows the potential of 3D imaging at high-resolution by phase-contrast tomography to better understand ossification in healing tendons.

Drug suspending during wound healing effectively weakens immunosuppression-related complications by preserving CD8+ T cell function.

Immunosuppressive medications, which interfere with the activation and proliferation of T and B cells, increase the risk of wound healing complications. To address it, this study aimed to validate the feasibility of drug suspending during wound healing, whilst exploring the mechanisms exerted by T cells, which are important in the wound healing process. For this, a mouse skin wound model was set up. Tacrolimus (FK506) and fingolimod (FTY720) were both administered intraperitoneally prior to wounding to inhibit the T cell activation and migration, respectively. Flow-cytometric analysis subsequently revealed the functional T cell subtypes detected during the healing process. A CD8a antibody was also administered to deplete CD8+ T cells in vivo to verify their specific function. It was found that FK506 or FTY720 administration delayed the early phase of wound healing by reducing collagen production, which was also supported by the downregulation of col1a1, col3a1 and tgfb1. However, there was no significant difference in the total healing period. Both spleen- and skin-derived CD8+ T cells were proliferated and activated after injury without intervention, whereas CD4+ T cells showed no significant changes. Furthermore, selectively depleting CD8+ T cells retarded the healing process by downregulating collagen production-associated genes (col1a1, col3a1, tgfβ1 and en1) and proteins (collagen type 1 and 3). In addition, the CD8a antibody decreased the expression of genes lta, tnfa, il13 and il13ra, and protein interleukin-13Rα. In conclusion, suspending immunosuppressive drugs during wound healing was shown to be feasible through restraining the migration of activated T cells. CD8+ T cells represented the primary functional subtype positively associated with wound healing.

Efektivitas Penggunaan Platelet-Rich Plasma (PRP) dalam Proses Penyembuhan Tulang pada Model Hewan Coba: Systematic Review

Bone injury causes thus, reduced productivity and death in the sufferer. Alternative treatment research was developed to speed up the bone healing process, including autologous treatments such as Platelet-Rich Plasma (PRP). Objective: To assessed the effectiveness of PRP use in the bone healing process on studies with experimental animal models. Methods: This study was a systematic review research whose article searches through the databases such as PubMed, ScienceDirect and ProQuest on English-language articles published from 2012-2022. The keywords used were ((bone healing OR fracture healing OR bone defect) AND (platelet rich plasma OR prp) AND (animal models)). Results: PRP was effective in accelerating bone regeneration in every experimental animal model in the early phases of healing, but not all studies have significant differences between groups that used PRP and those who did not. Conclusion: PRP has a positive impact in increasing bone mineral density, but the effectiveness of using PRP may vary from study result to study result, depending on the characteristics of the experimental animal model used in the study and the accuracy of concentration in the administration of PRP.Keywords: animal models, bone healing, platelet-rich plasma

Influence of Scaffold Microarchitecture on Angiogenesis and Regulation of Cell Differentiation during the Early Phase of Bone Healing: A Transcriptomics and Histological Analysis.

The early phase of bone healing is a complex and poorly understood process. With additive manufacturing, we can generate a specific and customizable library of bone substitutes to explore this phase. In this study, we produced tricalcium phosphate-based scaffolds with microarchitectures composed of filaments of 0.50 mm in diameter, named Fil050G, and 1.25 mm named Fil125G, respectively. The implants were removed after only 10 days in vivo followed by RNA sequencing (RNAseq) and histological analysis. RNAseq results revealed upregulation of adaptive immune response, regulation of cell adhesion, and cell migration-related genes in both of our two constructs. However, significant overexpression of genes linked to angiogenesis, regulation of cell differentiation, ossification, and bone development was observed solely in Fil050G scaffolds. Moreover, quantitative immunohistochemistry of structures positive for laminin revealed a significantly higher number of blood vessels in Fil050G samples. Furthermore, µCT detected a higher amount of mineralized tissue in Fil050G samples suggesting a superior osteoconductive potential. Hence, different filament diameters and distances in bone substitutes significantly influence angiogenesis and regulation of cell differentiation involved in the early phase of bone regeneration, which precedes osteoconductivity and bony bridging seen in later phases and as consequence, impacts the overall clinical outcome.

Ultrasonic-controlled “explosive” hydrogels to precisely regulate spatiotemporal osteoimmune disturbance

Spatiotemporal Immune disorder is a key factor leading to the failure of bone tissue healing. It is of vital importance to accurately suppress excessive peak immune response within 24-48 h of the injury and so regulate the spatiotemporal osteoimmune disturbance of bones. In this study, Ultrasound Controlled “Explosive” (UCE) hydrogels were prepared from gelatin-hyaluronic acid methacrylate hydrogels loaded with resveratrol nanobubbles produced by double emulsification through a condensation reaction. Such materials innovatively enable ultrasound-controlled RES release for precise regulation of spatiotemporal osteoimmune disorders. Under an ultrasonic power level of 1.5 W/cm2, the rate of effectively released RES through the blast of UCE hydrogels reached 38.14 %. And compared with the control group, the in vivo inhibition of inflammation and osteogenesis effects of UCE hydrogels were more effective, respectively. As suggested by the results, the excessive local inflammatory response was inhibited by the release of resveratrol, the temporospatial disorder of bone immune was precisely regulated, and as a result, the process of bone repair was accelerated. Altogether, this study confirms that the newly created UCE Hydrogels effectively promote bone repair by intervening peak inflammation during the early phase of fracture healing.

Altered early immune response after fracture and traumatic brain injury.

Clinical and preclinical data suggest accelerated bone fracture healing in subjects with an additional traumatic brain injury (TBI). Mechanistically, altered metabolism and neuro-endocrine regulations have been shown to influence bone formation after combined fracture and TBI, thereby increasing the bone content in the fracture callus. However, the early inflammatory response towards fracture and TBI has not been investigated in detail so far. This is of great importance, since the early inflammatory phase of fracture healing is known to be essential for the initiation of downstream regenerative processes for adequate fracture repair. Therefore, we analyzed systemic and local inflammatory mediators and immune cells in mice which were exposed to fracture only or fracture + TBI 6h and 24h after injury. We found a dysregulated systemic immune response and significantly fewer neutrophils and mast cells locally in the fracture hematoma. Further, local CXCL10 expression was significantly decreased in the animals with combined trauma, which correlated significantly with the reduced mast cell numbers. Since mast cells and mast cell-derived CXCL10 have been shown to increase osteoclastogenesis, the reduced mast cell numbers might contribute to higher bone content in the fracture callus of fracture + TBI mice due to decreased callus remodeling.

Microdialysis Reveals Anti-Inflammatory Effects of Sulfated Glycosaminoglycanes in the Early Phase of Bone Healing.

Although chronic inflammation inhibits bone healing, the healing process is initiated by an inflammatory phase. In a well-tuned sequence of molecular events, pro-inflammatory cytokines are secreted to orchestrate the inflammation response to injury and the recruitment of progenitor cells. These events in turn activate the secretion of anti-inflammatory signaling molecules and attract cells and mediators that antagonize the inflammation and initiate the repair phase. Sulfated glycosaminoglycanes (sGAG) are known to interact with cytokines, chemokines and growth factors and, thus, alter the availability, duration and impact of those mediators on the local molecular level. sGAG-coated polycaprolactone-co-lactide (PCL) scaffolds were inserted into critical-size femur defects in adult male Wistar rats. The femur was stabilized with a plate, and the defect was filled with either sGAG-containing PCL scaffolds or autologous bone (positive control). Wound fluid samples obtained by microdialysis were characterized regarding alterations of cytokine concentrations over the first 24 h after surgery. The analyses revealed the inhibition of the pro-inflammatory cytokines IL-1β and MIP-2 in the sGAG-treated groups compared to the positive control. A simultaneous increase of IL-6 and TNF-α indicated advanced regenerative capacity of sGAG, suggesting their potential to improve bone healing.