Violence in the Early Bronze Age. Diagnosis of skull lesions using anthropological, taphonomic and scanning electron microscopy techniques.

In this paper we present the study of a skull belonging to a young male from the Italian Bronze Age showing three perimortem injuries on the frontal and parietal bones; the peculiarity of the frontal injury is represented by its singular shape, which may be indicative of the weapon that caused the lesion. The aim of the present study is to examine the traumatic evidence in relation to possible etiological factors, in order to attempt to establish if the lesion occurred peri or post-mortem, and to evaluate if these traumatic injuries could be interpreted as an evidence of interpersonal violence, by combining anthropological, taphonomic and ESEM investigations. The combination of multidisciplinary methods of study can provide important new insights into inter-personal violence.

Photobiomodulation of mesenchymal stem cells encapsulated in an injectable rhBMP4-loaded hydrogel directs hard tissue bioengineering.

Photobiomodulation (PBM) therapy displays relevant properties for tissue healing and regeneration, which may be of interest for the tissue engineering field. Here, we show that PBM is able to improve cell survival and to interact with recombinant human Bone Morphogenetic Protein 4 (rhBMP4) to direct and accelerate odonto/osteogenic differentiation of dental derived mesenchymal stem cells (MSCs). MSCs were encapsulated in an injectable and thermo-responsive cell carrier (Pluronic® F-127) loaded with rhBMP4 and then photoactivated. PBM improved MSCs self-renewal and survival upon encapsulation in the Pluronic® F-127. In the presence of rhBMP4, cell odonto/osteogenic differentiation was premature and markedly improved in the photoactivated MSCs. An in vivo calvarial critical sized defect model demonstrated significant increase in bone formation after PBM treatment. Finally, a balance in the reactive oxygen species levels may be related to the favorable results of PBM and rhBMP4 association. PBM may act in synergism with rhBMP4 and is a promise candidate to direct and accelerate hard tissue bioengineering.

Histomorphometric and microtomographic evaluation of the effects of hyperbaric oxygen and systemic ozone, used alone and in combination, on calvarial defect healing in rats.

PURPOSE: The aim of this study was to compare the efficacy of hyperbaric oxygen and systemic ozone, used separately and in combination, on the healing of bone defects. MATERIAL AND METHODS: Sixty male Wistar rats were divided into 4 groups (n = 15) according to treatment (control, hyperbaric oxygen [HBO], ozone [O], and HBO plus O [HBO-O]) and divided further into 3 subgroups according to day of sacrifice (postsurgical days 5, 15, and 30). Surgery was performed under general anesthesia to create a critical-size bone defect (5 mm in diameter) in the cranium. After sacrifice, microtomographic images of all samples were recorded, and histomorphometric analysis was performed. RESULTS: Histologic and radiologic measurements showed that the values of all experimental groups were higher than those of the control group. Histologic scores for all experimental groups were statistically higher than those for the control group day 30 (O, P = .045; HBO, P = .049; HBO-O, P = .042). Histologic scores also were statistically higher for the HBO group on day 5 (P = .045) and day 15 (P = .009) compared with the control group. Microtomographic scores were higher for the experimental groups than for the control group, with statistically significant differences for group O on day 5 (P = .033) and day 30 (P = .0045) and for group HBO on day 15 (P = .005). Histologic and radiologic analyses showed positive correlations. CONCLUSION: Within the limitations of this study, the use of hyperbaric oxygen and ozone, separately and in combination, were shown to be effective in increasing bone healing. Combined usage was no more effective in stimulating bone healing than separate usage.

The effects of tissue-non-specific alkaline phosphatase gene therapy on craniosynostosis and craniofacial morphology in the FGFR2C342Y/+ mouse model of Crouzon craniosynostosis.

OBJECTIVES: Craniosynostosis, the premature fusion of cranial bones, has traditionally been described as a disease of increased bone mineralization. However, multiple mouse models of craniosynostosis display craniosynostosis simultaneously with diminished cranial bone volume and/or density. We propose an alternative hypothesis that craniosynostosis results from abnormal tissue mineralization through the downregulation of tissue-non-specific alkaline phosphatase (TNAP) enzyme downstream of activating mutations in FGFRs. MATERIAL AND METHODS: Neonatal Crouzon (FGFRC342Y/+) and wild-type (FGFR+/+) mice were injected with lentivirus to deliver a recombinant form of TNAP. Mice were sacrificed at 4 weeks postnatal. Serum was collected to test for alkaline phosphatase (AP), phosphorus, and calcium levels. Craniofacial bone fusion and morphology were assessed by micro-computed tomography. RESULTS: Injection with the TNAP lentivirus significantly increased serum AP levels (increased serum AP levels are indicative of efficient transduction and production of the recombinant protein), but results were variable and dependent upon viral lot and the litter of mice injected. Morphological analysis revealed craniofacial form differences for inferior surface (p=0.023) and cranial height (p=0.014) regions between TNAP lentivirus-injected and vehicle-injected Crouzon mice. With each unit increase in AP level, the odds of lambdoid suture fusion decreased by 84.2% and these results came close to statistical significance (p=0.068). CONCLUSION: These results suggest that TNAP deficiency may mediate FGFR2-associated craniosynostosis. Future studies should incorporate injection of recombinant TNAP protein, to avoid potential side effects and variable efficacy of lentiviral gene delivery.

Fatal cranial injury in an individual from Messina (Sicily) during the times of the Roman Empire.

Forensic and archaeological examinations of human skeletons can provide us with evidence of violence. In this paper, we present the patterns of two cranial lesions found on an adult male (T173) buried in a grave in the necropolis 'Isolato 96', Messina, Sicily, dating back to the Roman Empire (1st century BC - 1st century AD). The skull reveals two perimortem traumatic lesions, one produced by a sharp object on the right parietal bone and the other one on the left parietal bone, presumably the result of a fall. The interpretation of fracture patterns found in this cranium are an illustration of how forensic approaches can be applied with great benefit to archaeological specimens.

Effects of cigarette smoke inhalation and coffee consumption on bone formation and osseous integration of hydroxyapatite implant.

The present study aims to assess the effects of cigarette smoke inhalation and/or coffee consumption on bone formation and osseous integration of a dense hydroxyapatite (DHA) implant in rats. For this study, 20 male rats were divided into four groups (n = 5): CT (control) group, CE (coffee) group, CI (cigarette) group and CC (coffee + cigarette) group. During 16 weeks, animals in the CI group were exposed to cigarette smoke inhalation equivalent to 6 cigarettes per day; specimens in the CE group drank coffee as liquid diet; and rats in the CC group were submitted to both substances. In the 6th week a 5 mm slit in the parietal bone and a 4 mm slit in the tibia were performed on the left side: the former was left open while the latter received a DHA implant. As soon as surgeries were finished, the animals returned to their original protocols and after 10 weeks of exposure they were euthanised (ethically sacrificed) and the mentioned bones collected for histological processing. Data showed that exposure to cigarette smoke inhalation and coffee consumption did not interfere in weight gain and that solid and liquid diet consumption was satisfactory. Rats in the CC group showed a decrease in bone neoformation around the tibial DHA implant (31.8 ± 2.8) as well as in bone formation in the parietal slit (28.6 ± 2.2). On their own, cigarette smoke inhalation or coffee consumption also led to diminished bone neoformation around the implant and delayed the bone repair process in relation to the CT group. However, reduction in the bone repair process was accentuated with exposure to both cigarette smoke inhalation and coffee consumption in this study.

Effects of cigarette smoke inhalation and coffee consumption on bone formation and osseous integration of hydroxyapatite implant / Os efeitos do cigarro e do consumo de café sobre a formação óssea e a integração óssea de implantes de hidroxiapatita

The present study aims to assess the effects of cigarette smoke inhalation and/or coffee consumption on bone formation and osseous integration of a dense hydroxyapatite (DHA) implant in rats. For this study, 20 male rats were divided into four groups (n = 5): CT (control) group, CE (coffee) group, CI (cigarette) group and CC (coffee + cigarette) group. During 16 weeks, animals in the CI group were exposed to cigarette smoke inhalation equivalent to 6 cigarettes per day; specimens in the CE group drank coffee as liquid diet; and rats in the CC group were submitted to both substances. In the 6th week a 5 mm slit in the parietal bone and a 4 mm slit in the tibia were performed on the left side: the former was left open while the latter received a DHA implant. As soon as surgeries were finished, the animals returned to their original protocols and after 10 weeks of exposure they were euthanised (ethically sacrificed) and the mentioned bones collected for histological processing. Data showed that exposure to cigarette smoke inhalation and coffee consumption did not interfere in weight gain and that solid and liquid diet consumption was satisfactory. Rats in the CC group showed a decrease in bone neoformation around the tibial DHA implant (31.8 ± 2.8) as well as in bone formation in the parietal slit (28.6 ± 2.2). On their own, cigarette smoke inhalation or coffee consumption also led to diminished bone neoformation around the implant and delayed the bone repair process in relation to the CT group. However, reduction in the bone repair process was accentuated with exposure to both cigarette smoke inhalation and coffee consumption in this study.

O presente estudo teve como objetivo avaliar os efeitos do tabagismo e do consumo de café, isolada ou concomitantemente, sobre a formação óssea e a osseointegração de implantes hidroxiapatita densa. Foram utilizados 20 ratos machos, divididos em quatro grupos (n = 5): grupo CT (controle); grupo CA (café); grupo CI (cigarro), e grupo CC (cigarro + café). Durante 16 semanas, os animais do grupo CI foram expostos à fumaça de seis cigarros/dia; os animais do grupo CA consumiram café como dieta líquida, e os animais do grupo CC, ambas as substâncias. Após seis semanas de exposição, uma falha óssea de 5 mm foi produzida no osso parietal esquerdo e de 4 mm, na tíbia esquerda dos animais. A falha do parietal foi mantida aberta, enquanto na tíbia corpos cerâmicos de hidroxiapatita densa (HAD) foram implantados em cavidade produzida cirurgicamente. Após as cirurgias, os animais retornaram aos protocolos experimentais e, ao término de dez semanas, foram eutanasiados, sendo as tíbias e os parietais coletados para processamento histológico. A exposição à fumaça do cigarro e o consumo de café não interferiram no ganho de peso dos animais, e os consumos de dieta líquida e sólida foram satisfatórios entre os grupos. Os animais do grupo CC apresentaram menor volume de osso neoformado ao redor do implante de HAD na tíbia (31,8 ± 2,8) e menor osteogênese na falha produzida no osso parietal (28,6 ± 2,2). O café e o cigarro consumidos isoladamente provocam a diminuição do volume de osso ao redor do implante e o atraso no processo de reparação óssea. Observou-se que o consumo de café associado à exposição à fumaça do cigarro reduziu de forma acentuada o processo de reparação óssea, no presente estudo.

Effect of Buguzhi (Psoralea corylifolia fruit) extract on bone formation.

The objective of the study is to compare the amount of new bone produced by Buguzhi (Psoralea corylifolia fruit) extract in collagen matrix to that produced and collagen matrix in vivo. Eighteen bone defects, 5 mm by 10 mm, were created in the parietal bone of 9 New Zealand white rabbits. Six defects were grafted with Buguzhi extract mixed with collagen matrix. Six defects were grafted with collagen matrix alone (positive control) and 6 were left empty (negative control). Animals were sacrificed on day 14 and the defects were dissected and prepared for histological assessment. Quantitative analysis of new bone formation and bone cells was made on 100 sections (50 sections for each group) using image analysis. A total of 275% more new bone was present in defects grafted with Buguzhi extract in collagen matrix than those grafted with collagen matrix. No bone was formed in the negative control group. The amount of bone cells was also significantly greater in the Buguzhi group than in the positive control group. To conclude, Buguzhi extract in collagen matrix has the effect of increasing new bone formation locally in vivo. Buguzhi extract in collagen matrix can be used as a bone graft material.

Long-term histologic analysis of bone tissue alteration and healing following Er:YAG laser irradiation compared to electrosurgery.

BACKGROUND: The erbium-doped:yttrium, aluminum, and garnet (Er:YAG) laser is reportedly useful for periodontal therapy. However, the potential thermal damage that Er:YAG laser irradiation can produce on bone tissue has not been fully clarified. The purpose of this study was to histologically examine the effects of the Er:YAG laser on bone tissue and subsequent wound healing compared to electrosurgery in a long-term study. METHODS: Calvarial bone from 30 rats was exposed to contact and non-contact Er:YAG laser irradiation (115 mJ/pulse, 10 Hz) without water coolant, or electrode contact. The treated surfaces were analyzed by scanning electron microscopy (SEM), and the healing process was histologically observed until 12 months post-surgery. RESULTS: Contact irradiation resulted in substantial bone ablation, whereas non-contact irradiation produced slight tissue removal. Histologic and SEM analyses of the lased surface showed no severe thermal damage, except for the production of a superficially affected layer with a microstructured surface. The layer did not inhibit new bone formation, and the ablated defect was repaired uneventfully. Although the thickness of the layer gradually decreased, it generally remained in the cortical bone through the observation period. Electrosurgery produced a large area of thermal necrosis without ablation, and the damaged area was not replaced with new bone. CONCLUSIONS: Unlike electrosurgery, Er:YAG laser irradiation without water coolant easily ablated bone tissue, and thermal alteration in the treated surface was minimal. The superficially affected layer did not interfere with the ensuing bone healing, resulting in favorable repair of the defect.

Effect of Salvia miltiorrhiza extract on bone formation.

Salvia miltiorrhiza (SM) is a Chinese medicine used for treatment of cardiac diseases through improving blood circulation. Our laboratory compared the amount of new bone formed in collagen matrix with SM extract to that formed in the collagen matrix alone. Eighteen bone defects, 5 mm x 10 mm were created in the parietal bone of nine New Zealand White rabbits. In the experimental group, six defects were grafted with collagen matrix with SM extract. In the control groups, six defects were grafted with collagen matrix alone (positive control) and six were left empty (negative control). Animals were killed on day 14 and the defects were dissected and prepared for histological assessment. Serial sections were cut across each defect. Quantitative analysis of new bone formation was made on 100 sections (50 sections for each group of SM and positive control) using image analysis. A total of 478% more new bone was present in defects grafted with SM extract in collagen matrix than those grafted with the collagen matrix alone. Very little new bone was formed in the passive control group. In conclusion, SM extract in collagen matrix has the effect of increasing new bone formation locally and can be used for bone grafting especially in cases with compromised vascular responses.

Effect of puerarin on bone formation.

OBJECTIVE: Puerarin is one of the major phytoestrogens isolated from Pueraria lobata, a Chinese medicine known as Gegen. Our laboratory compared the amount of new bone produced by puerarin in collagen matrix (carrier) to that produced by the collagen matrix alone. METHOD: Eighteen bone defects, 5mm by 10mm were created in the parietal bone of nine New Zealand White rabbits. In the experimental group, six defects were grafted with puerarin solution mixed with collagen matrix. In the control groups, six defects were grafted with collagen matrix alone (active control) and six were left empty (passive control). Animals were killed on day 14 and the defects were dissected and prepared for histological assessment. Serial sections were cut across each defect. No new bone was formed in the passive control group. Quantitative analysis of new bone formation was made on 100 sections (10 sections in each defect, in five defects randomly selected in each of the experimental group and active control group) using image analysis. RESULTS: A total of 554% more new bone was present in defects grafted with puerarin in collagen matrix than those grafted with the collagen matrix alone. CONCLUSION: Puerarin in collagen matrix has the effect of increasing new bone formation locally and can be used for bone grafting or for bone induction often required in surgery.

Thyroid follicular carcinoma metastasized to the lung, skull, and brain 12 years after initial treatment for thyroid gland--case report.

A 65-year-old woman presented with multiple metastases from thyroid follicular carcinoma to the lung, skull, and brain. The skull and brain tumors had been successfully treated by surgery, thyroxine supplementation, and radiosurgery until she died of sudden intracerebral hemorrhage which had no connection with tumor treatment. The lung tumor was treated by conventional irradiation and radioactive ablation. Well-differentiated thyroid carcinoma is a slowly progressive tumor. Follicular carcinoma is thought to have the most optimistic prognosis even with metastases to the lymph nodes and lung. Radioactive ablation using iodine-131 is widely used to treat the primary and/or metastatic lesion. However, the prognosis for patients with brain metastases is poor. Intracranial metastasis of this tumor is rare, but has a mean posttreatment survival of around 12 months. Surgical excision of the metastatic intracranial lesion may be the only effective treatment.

Hyperbaric oxygen results in an increase in rabbit calvarial critical sized defects.

OBJECTIVE: This study was undertaken to evaluate whether the effects of hyperbaric oxygen (HBO) therapy could alter the critical size for spontaneous healing of a bone defect in the rabbit calvarial model. STUDY DESIGN: An animal trial of 12 weeks duration was conducted using 20 New Zealand white rabbits, which were randomly divided into 2 groups of 10 animals each. Calvarial defects were created in the parietal bones of each animal bilaterally. Defects were critical-sized, 15 mm on one side and supra-critical-sized, 18 mm on the contralateral side. Group 1 received a 90-min HBO treatment sessions at 2.4 absolute atmospheric pressure (ATA) per day for 20 consecutive days. Group 2 served as a control without any HBO treatment sessions. Five animals in each group were sacrificed at 6 and 12 weeks. Data analysis included qualitative assessment of the calvarial specimens, post-sacrifice radiographs, as well as histomorphometric analysis to compute the amount of regenerated bone within the defects. ANOVA and paired sample t test were used for statistical analysis. RESULTS: Both radiographic analysis and histomorphometric analysis demonstrated that HBO-treated animals had significantly more new bone within their defects compared with the control group (P < .001). There was no statistically significant difference between the percentage of new bone forming in the 15-mm and 18-mm HBO-treated defects. There was no difference between the 6-week and the 12-week HBO-treated groups. HBO is effective in enhancing the bony healing of full thickness critical sized as well as supra-critical-sized defects in the rabbit calvarial model. CONCLUSION: Bone regeneration was significantly greater in the HBO-treated animals regardless of the defect size. HBO may have increased the diameter of the rabbit critical-sized calvarial defect to more than 18 mm.

Histological and TEM examination of early stages of bone healing after Er:YAG laser irradiation.

OBJECTIVE: The aim of this study was to analyze the early healing process of bone tissue irradiated by Er:YAG laser and compare it with that treated by mechanical drilling and CO(2) laser. BACKGROUND DATA: Er:YAG laser has a great potential for cutting hard tissues as it is capable of ablation with less thermal damage. METHODS: Twenty-four male Wistar rats were used for this study. The calvarial bone of rats was exposed and straight grooves were prepared by Er:YAG laser, mechanical bur and continuous wave CO(2) laser. Four rats each were sacrificed at six time points: 10 min, 6 and 24 h and 3, 7, and 14 days post-surgery. Sections were prepared for light and transmission electron microscopic (TEM) observations. RESULTS: Compared to mechanical bur and CO(2) groups, the inflammatory cell infiltration adjacent to the irradiated bone surface, fibroblastic reaction, and revascularization were more pronounced in the Er:YAG laser-irradiated tissues. A cell-rich granulation tissue with fibroblasts and osteoblasts was predominant in 7-day specimens of Er:YAG laser group. Histopathological analysis of 14-day specimens in the Er:YAG group also revealed significantly greater new bone formation, compared with the mechanical bur and CO(2) laser groups. CONCLUSIONS: Initial bone healing following Er:YAG laser irradiation occurred faster than that after mechanical bur and CO(2) laser. Er:YAG laser treatment may be advantageous for wound healing of bone tissue, presumably by providing a favorable surface for cell attachment.

Enhancement of bone formation in rat calvarial bone defects using low-level laser therapy.

OBJECTIVE: To evaluate the effect of low-level laser therapy (LLLT), using a GaAlAs diode laser device, on bone healing and growth in rat calvarial bone defects. STUDY DESIGN: An animal trial of 4 weeks' duration was conducted using a randomized blind, placebo-controlled design. Standardized round osseous defects of 2.7 mm diameter were made in each parietal bone of 20 rats (n=40 defects). The animals were randomly divided into an experimental and a control group of 10 animals each. In the experimental group, a GaAlAs diode laser was applied immediately after surgery and then daily for 6 consecutive days. The control group received the same handling and treatment, but with the laser turned off. Five rats from each group were killed on day 14 and the remainder on day 28 postoperatively. From each animal, tissue samples from one defect were prepared for histochemistry and samples from the contralateral defect for histology. Levels of calcium, phosphorus, and protein were determined by using atomic absorption spectrometry, colorimetry, and photometry, respectively. Student t-test and Mann-Whitney were used for statistical analyses. RESULTS: At both time points the tissue samples from the experimental animals contained significantly more calcium, phosphorus, and protein than the controls. Similarly, histological analyses disclosed more pronounced angiogenesis and connective tissue formation, and more advanced bone formation in the experimental group than in the controls. CONCLUSION: LLLT may enhance bone formation in rat calvarial bone defects.

Brachycephaly and external pressure.

Browsing through some literatures, I happened to find out some records on the rite of imitation of the child's head to brachycephaly. In 1976, two skulls showing extraordinary forms were excavated out of an ancient tomb at Yean village of Daedong township in Kimhae County in Kyungnam Province, the Republic of Korea. The deformation in Yean village is judged to be an intentional frontal flattening. The frontal bone of deformed individuals is remarkably flattened by a primary deforming force. The parietal bone expands laterally and superoposteriorly as a major compensatory growth. The lambdoid or occipital flattening induced by the counterforce is light and unstable. In the facial skeleton, the maxilla shifts downward as a minor compensatory growth. These findings might be substantially correspondent to the custom in the southern area of Korea in the 4th century A.D. paragraphed by Han chuan of Wei-dynasty record, History of the Three Kingdoms. It described that the people gave a pressure onto the child's forehead with stones in order to mold it for preference. Flatheaded meant "free" among the Chinook and Salish tribes of American Indians. Babies of slaves were denied the year-long beauty treatment--a board tied to the infants' padded skull--that permanently flattened the foreheads of tribal members. Referring to these records, I presume that brachycephaly can be induced by direct extrinsic pressure at least until the soft infant's head gets hard.