Giuseppe Intini

The use of platelet-rich plasma in bone reconstruction therapy.

The use of platelet-rich plasma (PRP) in bone reconstruction therapy was introduced in the late 1990s. Since then, many scientists and clinicians have employed it in orthopaedic and oral surgeries. Unfortunately, studies that analyze the use of PRP are somewhat controversial as some conclude that the use of PRP may favor bone regeneration and others conclude that the use of PRP is irrelevant. By listing and analyzing the biological effect that each factor released by the activated platelets can have in bone regeneration, the present review answers the question of why PRP may be useful in bone reconstruction therapy. Subsequently, by examining the studies that have both successfully and unsuccessfully utilized PRP, it suggests how PRP might be used in order to achieve successful results in orthopaedic and dental bone reconstruction surgeries.

Calcium sulfate and platelet-rich plasma make a novel osteoinductive biomaterial for bone regeneration.

BackgroundWith the present study we introduce a novel and simple biomaterial able to induce regeneration of bone. We theorized that nourishing a bone defect with calcium and with a large amount of activated platelets may initiate a series of biological processes that culminate in bone regeneration. Thus, we engineered CS-Platelet, a biomaterial based on the combination of Calcium Sulfate and Platelet-Rich Plasma in which Calcium Sulfate also acts as an activator of the platelets, therefore avoiding the need to activate the platelets with an agonist.MethodsFirst, we tested CS-Platelet in heterotopic (muscle) and orthotopic (bone) bone regeneration bioassays. We then utilized CS-Platelet in a variety of dental and craniofacial clinical cases, where regeneration of bone was needed.ResultsThe heterotopic bioassay showed formation of bone within the muscular tissue at the site of the implantation of CS-Platelet. Results of a quantitative orthotopic bioassay based on the rat calvaria critical size defect showed that only CS-Platelet and recombinant human BMP2 were able to induce a significant regeneration of bone. A non-human primate orthotopic bioassay also showed that CS-Platelet is completely resorbable. In all human clinical cases where CS-Platelet was used, a complete bone repair was achieved.ConclusionThis study showed that CS-Platelet is a novel biomaterial able to induce formation of bone in heterotopic and orthotopic sites, in orthotopic critical size bone defects, and in various clinical situations. The discovery of CS-Platelet may represent a cost-effective breakthrough in bone regenerative therapy and an alternative or an adjuvant to the current treatments.

Periosteum Biology and Applications in Craniofacial Bone Regeneration

The bone-regenerative potentials of the periosteum have been explored as early as the 17th century. Over the past few years, however, much has been discovered in terms of the molecular and cellular mechanisms that control the periosteal contribution to bone regeneration. Lineage tracing analyses and knock-in transgenic mice have helped define the relative contributions of the periosteum and endosteum to bone regeneration. Additional studies have shed light on the critical roles that BMP, FGF, Hedgehog, Notch, PDGF, Wnt, and inflammation signaling have or may have in periosteal-mediated bone regeneration, fostering the path to novel approaches in bone-regenerative therapy. Thus, by examining the role that each pathway has in periosteal-mediated bone regeneration, in this review we analyze the status of the current research on the regenerative potential of the periosteum. The provided analysis aims to inform both clinician-scientists who may have interest in the current studies about the biology of the periosteum as well as dental surgeons who may find this review useful to perform periosteal-harnessing bone-regenerative procedures.The bone-regenerative potentials of the periosteum have been explored as early as the 17th century. Over the past few years, however, much has been discovered in terms of the molecular and cellular mechanisms that control the periosteal contribution to bone regeneration. Lineage tracing analyses and knock-in transgenic mice have helped define the relative contributions of the periosteum and endosteum to bone regeneration. Additional studies have shed light on the critical roles that BMP, FGF, Hedgehog, Notch, PDGF, Wnt, and inflammation signaling have or may have in periosteal-mediated bone regeneration, fostering the path to novel approaches in bone-regenerative therapy. Thus, by examining the role that each pathway has in periosteal-mediated bone regeneration, in this review we analyze the status of the current research on the regenerative potential of the periosteum. The provided analysis aims to inform both clinician-scientists who may have interest in the current studies about the biology of the periosteum as well as dental surgeons who may find this review useful to perform periosteal-harnessing bone-regenerative procedures.

Engineering a Bioactive Matrix by Modifications of Calcium Sulfate

The goal of this study was to define the conditions for the fabrication of a bioactive matrix that induces and supports cell proliferation and tissue regeneration. The proposed hypothesis was that a composite graft could be engineered by the absorption of platelet-rich plasma (PRP) onto calcium sulfate (CS). Evaluation of the biological activity of the engineered grafts was based on osteoblast proliferation studies and scanning electron microscopy (SEM) analyses. Graft samples were created in a standard size and shape so that the surface available for attachment and cell proliferation was always identical. Proliferation data were expressed as counts per minute per group and differences among groups were statistically analyzed by analysis of variance followed by the Scheffé test (alpha = 0.1). SEM analysis showed that the combination of CS and PRP presents a preserved crystalline structure well integrated by organic matrix. This combination showed the highest cell proliferation levels (p < 0.001). Further evaluations demonstrated that PRP is activated when combined with CS. When tested as a possible carrier for biologically active molecules such as platelet-derived growth factor (PDGF), CS showed increased cell proliferation (p < 0.001). SEM revealed adherent osteoblasts with broad flattened edges on CS-PRP. This study proposes CS as an efficient carrier for PRP or PDGF and supports the use of these combinations as bioactive matrices in clinical or laboratory applications.

Potential for Stem Cell‐Based Periodontal Therapy

Periodontal diseases are highly prevalent and are linked to several systemic diseases. The goal of periodontal treatment is to halt the progression of the disease and regenerate the damaged tissue. However, achieving complete and functional periodontal regeneration is challenging because the periodontium is a complex apparatus composed of different tissues, including bone, cementum, and periodontal ligament. Stem cells may represent an effective therapeutic tool for periodontal regeneration due to their plasticity and their ability to regenerate different tissues. This review presents and critically analyzes the available information on stem cell‐based therapy for the regeneration of periodontal tissues and suggests new avenues for the development of more effective therapeutic protocols. J. Cell. Physiol. 230: 50–61, 2016.

Future Approaches in Periodontal Regeneration: Gene Therapy, Stem Cells, and RNA Interference

Periodontal disease is a major public health issue and the development of effective therapies to treat the disease and regenerate periodontal tissue is an important goal of todays medicine. This article highlights recent scientific advancements in gene therapy, stem cell biology, and RNA interference with the intent of identifying their potential in periodontal tissue regeneration. Results from basic research, preclinical, and clinical studies indicate that these fields of research may soon contribute to more effective regenerative therapies for periodontal disease.

Femtosecond laser bone ablation with a high repetition rate fiber laser source

Femtosecond laser pulses can be used to perform very precise cutting of material, including biological samples from subcellular organelles to large areas of bone, through plasma-mediated ablation. The use of a kilohertz regenerative amplifier is usually needed to obtain the pulse energy required for ablation. This work investigates a 5 megahertz compact fiber laser for near-video rate imaging and ablation in bone. After optimization of ablation efficiency and reduction in autofluorescence, the system is demonstrated for the in vivo study of bone regeneration. Image-guided creation of a bone defect and longitudinal evaluation of cellular injury response in the defect provides insight into the bone regeneration process.

Postnatal Calvarial Skeletal Stem Cells Expressing PRX1 Reside Exclusively in the Calvarial Sutures and Are Required for Bone Regeneration

Summary Post-natal skeletal stem cells expressing PRX1 (pnPRX1+) have been identified in the calvaria and in the axial skeleton. Here we characterize the location and functional capacity of the calvarial pnPRX1+ cells. We found that pnPRX1+ reside exclusively in the calvarial suture niche and decrease in number with age. They are distinct from preosteoblasts and osteoblasts of the sutures, respond to WNT signaling in vitro and in vivo by differentiating into osteoblasts, and, upon heterotopic transplantation, are able to regenerate bone. Diphtheria toxin A (DTA)-mediated lineage ablation of pnPRX1+ cells and suturectomy perturb regeneration of calvarial bone defects and confirm that pnPRX1+ cells of the sutures are required for bone regeneration. Orthotopic transplantation of sutures with traceable pnPRX1+ cells into wild-type animals shows that pnPRX1+ cells of the suture contribute to calvarial bone defect regeneration. DTA-mediated lineage ablation of pnPRX1+ does not, however, interfere with calvarial development.

Efficacy of human salivary mucin MUC7-derived peptide and histatin 5 in a murine model of candidiasis

MUC7 16-mer (residues 36-51 of human salivary mucin, MUC7) and histatin 5 possess potent in vitro antifungal activity. In the present study, we have evaluated the efficacy of these peptides in vivo using the experimental model of murine vulvo-vaginal candidiasis. The treatment groups included MUC7 16-mer, histatin 5, clotrimazole (all in pluronic F127 gel), and placebo (gel alone). Mice were treated intravaginally for 7 consecutive days. At the end of the treatment, anticandidal activities were assessed by colony counts and by histological examination. All groups except clotrimazole presented positive cultures; no statistically significant differences were found in fungal burden amongst placebo and any treatment group except clotrimazole. Histopathological findings confirmed the microbiological results; all groups with the exception of clotrimazole showed variable signs of infection.

Alveolar Bone Loss Mechanisms, Potential Therapeutic Targets, and Interventions

This article reviews recent research into mechanisms underlying bone resorption and highlights avenues of investigation that may generate new therapies to combat alveolar bone loss in periodontitis. Several proteins, signaling pathways, stem cells, and dietary supplements are discussed as they relate to peri-odontal bone loss and regeneration. RGS12 is a crucial protein that mediates osteoclastogenesis and bone destruction, and a potential therapeutic target. RGS12 likely regulates osteoclast differentiation through regulating calcium influx to control the calcium oscillation-NFATc1 pathway. A working model for RGS10 and RGS12 in the regulation of Ca2+ oscillations during osteoclast differentiation is proposed. Initiation of inflammation depends on host cell-microbe interactions, including the p38 mitogen-activated protein kinase (MAPK) signaling pathway. Oral p38 inhibitors reduced lipopolysaccharide (LPS)-induced bone destruction in a rat periodontitis model but showed unsatisfactory safety profiles. The p38 substrate MK2 is a more specific therapeutic target with potentially superior tolerability. Furthermore, MKP-1 shows anti-inflammatory activity, reducing inflammatory cytokine biosynthesis and bone resorption. Multipotent skeletal stem cell (SSC) populations exist within the bone marrow and periosteum of long bones. These bone-marrow-derived SSCs and periosteum-derived SSCs have shown therapeutic potential in several applications, including bone and periodontal regeneration. The existence of craniofacial bone-specific SSCs is suggested based on existing studies. The effects of calcium, vitamin D, and soy isoflavone supplementation on alveolar and skeletal bone loss in post-menopausal women were investigated. Supplementation resulted in stabilization of forearm bone mass density and a reduced rate of alveolar bone loss over 1 yr, compared with placebo. Periodontal attachment levels were also well-maintained and alveolar bone loss suppressed during 24 wk of supplementation.