Novel Therapeutic Effects in Rat Spinal Cord Injuries: Recovery of the Definitive and Early Spinal Cord Injury by the Administration of Pentadecapeptide BPC 157 Therapy.

Recently, marked therapeutic effects pertaining to the recovery of injured rat spinal cords (1 min compression injury of the sacrocaudal spinal cord (S2-Co1) resulting in tail paralysis) appeared after a single intraperitoneal administration of the stable gastric pentadecapeptide BPC 157 at 10 min post-injury. Besides the demonstrated rapid and sustained recovery (1 year), we showed the particular points of the immediate effect of the BPC 157 therapy that began rapidly after its administration, (i) soon after injury (10 min), or (ii) later (4 days), in the rats with a definitive spinal cord injury. Specifically, in counteracting spinal cord hematoma and swelling, (i) in rats that had undergone acute spinal cord injury, followed by intraperitoneal BPC 157 application at 10 min, we focused on the first 10-30 min post-injury period (assessment of gross, microscopic, and gene expression changes). Taking day 4 post-injury as the definitive injury, (ii) we focused on the immediate effects after the BPC 157 intragastric application over 20 min of the post-therapy period. Comparable long-time recovery was noted in treated rats which had definitive tail paralysis: (iii) the therapy was continuously given per orally in drinking water, beginning at day 4 after injury and lasting one month after injury. BPC 157 rats presented only discrete edema and minimal hemorrhage and increased Nos1, Nos2, and Nos3 values (30 min post-injury, (i)) or only mild hemorrhage, and only discrete vacuolation of tissue (day 4, (ii)). In the day 4-30 post-injury study (iii), BPC 157 rats rapidly presented tail function recovery, and no demyelination process (Luxol fast blue staining).

Pentadecapeptide BPC 157 and the central nervous system.

We reviewed the pleiotropic beneficial effects of the stable gastric pentadecapeptide BPC 157, three very recent demonstrations that may be essential in the gut-brain and brain-gut axis operation, and therapy application in the central nervous system disorders, in particular. Firstly, given in the reperfusion, BPC 157 counteracted bilateral clamping of the common carotid arteries-induced stroke, sustained brain neuronal damages were resolved in rats as well as disturbed memory, locomotion, and coordination. This therapy effect supports particular gene expression in hippocampal tissues that appeared in BPC 157-treated rats. Secondly, there are L-NG-nitro arginine methyl ester (L-NAME)- and haloperidol-induced catalepsy as well as the rat acute and chronic models of 'positive-like' schizophrenia symptoms, that BPC 157 counteracted, and resolved the complex relationship of the nitric oxide-system with amphetamine and apomorphine (dopamine agents application), MK-801 (non-competitive antagonist of the N-methyl-D-aspartate receptor) and chronic methamphetamine administration (to induce sensitivity). Thirdly, after rat spinal cord compression, there were advanced healing and functional recovery (counteracted tail paralysis). Likewise, in BPC 157 therapy, there is specific support for each of these topics: counteracted encephalopathies; alleviated vascular occlusion disturbances (stroke); counteracted dopamine disturbances (dopamine receptors blockade, receptors super sensitivity development, or receptor activation, over-release, nigrostriatal damage, vesicles depletion), and nitric oxide-system disturbances ("L-NAME non-responsive, L-arginine responsive," and "L-NAME responsive, L-arginine responsive") (schizophrenia therapy); inflammation reduction, nerve recovery in addition to alleviated hemostasis and vessels function after compression (spinal cord injury therapy). Thus, these disturbances may be all resolved within the same agent's beneficial activity, i.e., the stable gastric pentadecapeptide BPC 157.

Stable gastric pentadecapeptide BPC 157 can improve the healing course of spinal cord injury and lead to functional recovery in rats.

BACKGROUND: We focused on the therapeutic effects of the stable gastric pentadecapeptide BPC 157 in spinal cord injury using a rat model. BPC 157, of which the LD1 has not been achieved, has been implemented as an anti-ulcer peptide in inflammatory bowel disease trials and recently in a multiple sclerosis trial. In animals, BPC 157 has an anti-inflammatory effect and therapeutic effects in functional recovery and the rescue of somatosensory neurons in the sciatic nerve after transection, upon brain injury after concussive trauma, and in severe encephalopathies. Additionally, BPC 157 affects various molecular pathways. METHODS: Therefore, BPC 157 therapy was administered by a one-time intraperitoneal injection (BPC 157 (200 or 2 µg/kg) or 0.9% NaCl (5 ml/kg)) 10 min after injury. The injury procedure involved laminectomy (level L2-L3) and a 60-s compression (neurosurgical piston (60-66 g) of the exposed dural sac of the sacrocaudal spinal cord). Assessments were performed at 1, 4, 7, 15, 30, 90, 180, and 360 days after injury. RESULTS: All of the injured rats that received BPC 157 exhibited consistent clinical improvement, increasingly better motor function of the tail, no autotomy, and resolved spasticity by day 15. BPC 157 application largely counteracted changes at the microscopic level, including the formation of vacuoles and the loss of axons in the white matter, the formation of edema and the loss of motoneurons in the gray matter, and a decreased number of large myelinated axons in the rat caudal nerve from day 7. EMG recordings showed a markedly lower motor unit potential in the tail muscle. CONCLUSION: Axonal and neuronal necrosis, demyelination, and cyst formation were counteracted. The functional rescue provided by BPC 157 after spinal cord injury implies that BPC 157 therapy can impact all stages of the secondary injury phase.

Radiopaque Gelified Ethanol Application in Lumbar Intervertebral Soft Disc Herniations: Croatian Multicentric Study.

Objective: Minimally invasive percutaneous spinal procedures are popular in trying to reduce spinal pain. The aim of this paper is to evaluate the safety of intervertebral disc chemonucleolysis and to report the effectiveness of a percutaneous, minimally invasive treatment for contained herniated intervertebral discs in the lumbar spine using the recently marketed radiopaque gelified ethanol. Methods: Pain relief before and after the procedure was self-evaluated by each patient using a verbal numeric scale (VNS) ranging from 0 to 10. Patients were also scored prior to procedure and after chemonucleolysis during several follow-up periods using the Roland-Morris low back pain and disability questionnaire (RMQ). Follow-up periods were defined as 0-6, 6-12, 12-18, 18-24, and 24-30 months. Clinically significant functional improvement (CSFI) was defined as a decrease of five or more points on the RMQ scale and a decrease of at least 50% of pain intensity using VNS. Results: Using the RMQ scale, CSFI was achieved in 20/29 patients in the first follow-up period, 20/27 patients in the second follow-up period, 9/12 patients in the third follow-up period, 8/9 patients in the fourth follow-up period, and 4/4 patients in the last follow-up period. Using the VNS rating, CSFI was accomplished in 19/29 patients in the first follow-up period, 19/27 patients in the second follow-up period, 9/12 patients in the third follow-up period, 8/9 patients in the fourth follow-up period, and 4/4 in the last follow-up period. Conclusions: Intradiscal application of gelified ethanol may be effective in pain reduction using the VNS and Roland-Morris low back pain and disability questionnaire. The treatment is safe and easy to handle.

[Diagnostics and treatment of osteoporotic vertebral fractures]. / Dijagnostika i lijecenje osteoporotskih prijeloma kraljezaka.

Osteoporosis is a systemic skeletal disease characterized by low bone mass and microarchitectural deterioration of bone tissue, with vertebral compressive fractures (VCFs) that may occur as a consequence. Vertebral compression fractures (VCFs) can lead to severe acute and chronic pain, impaired mobility, reduced quality of life and an increased risk of mortality due to decreased mobility and pulmonary dysfunction. When painful VCFs do come to clinical attention, they are typically treated with optimal pain management (OPM). Although the natural course of pain due to vertebral fractures decreases within the first weeks in the majority of patients, a number of them remain with persistent pain and/or ongoing vertebral collapse. Facet joint injections are an emerging procedure as possible treatment for a subgroup of patients with persistent pain after VCFs. In case of vertebral collapse or persistent pain after facet joint injections, patients have to be treated with percutaneous vertebroplasty (PVP) or percutaneous balloon kyphoplasty (BKP). PVP is a minimally invasive surgical procedure in which bone cement is injected into a fractured vertebra under radiological guidance using fluoroscopy. BKP is a variation of this approach, in which an inflatable balloon tamp is placed in the collapsed vertebra prior to cement injection, in order to create a cavity allowing low pressure injection. For peo- ple with painful osteoporotic VCFs refractory to analgesic treatment, PVP and BKP perform significantly better in unblinded trials than OPM in terms of improving quality of life and reducing pain and disability. It is possible that BKP and PVP may lead to reductions in mortality. In small subgroup of patients with neurological injury related to an osteoporotic fracture, different open surgical techniques were used to suit different fracture patterns, with good clinical and radiological results.

[First guidelines of Croatian interest group in diagnosing and treating pain conditions of cervical and thoracic spine using minimally invasive procedures]. / Prve hrvatske smjernice za dijagnostiku i lijecenje bolnih stanja vratne i prsne kraljeznice minimalno invazivnim postupcima.

Pain syndromes originating from cervical and thoracic spine remain to be a major public health problem. Medical expenses in general and surgical procedures associated with overall care for the neck and thoracic pain are high and growing. Furthermore, these two chronic pain conditions are also leading causes for missed workdays. Chronic pain syndromes originating from cervical spine are most commonly caused by degenerative changes of the facet joints. Cervi- cobrachial syndrome is most commonly caused by herniated discs. Diagnostic controlled blocks, performed in order to identify, the source of pain, often predetermine patient for further therapeutic minimally invasive interventions. If the chronic pain syndromes of the cervical and thoracic spine are caused by degenerative facet joints, patient can be offered neuroablative procedures using radiofrequency. In patients suffering from chronic cervical and thoracic pain caused by painful intervertebral disc minimally invasive intradiscal decompression procedures can be performed. In cases where the neck pain and radicular pain are caused by the central and foraminal spinal stenosis patients are advised epidural steroid injections. The purpose of above advised procedures, using steroids, local anesthetics and RF current, is to relieve patients' pain, allow optimal physical therapy, and improved functional capacity, consequently providing a better quality of life.

[First guidelines of Croatian interest group in diagnosing and treating lower back and radicular pain using minimally invasive diagnostic and therapeutic procedures]. / Prve hrvatske smjernice za dijagnostiku i lijecenje krizobolje i lumboishialgije minimalno invazivnim procedurama.

Low back and radicular pain recently became a major public health problem. Medical expenses in general, and surgical procedures associated with overall care for the lower back pain are high and growing. Furthermore, these two chronic pain conditions are also leading causes for missed workdays. Degenerative changes of the intervertebral disc, facet joints, sacroiliac joint or disc herniation as described during imaging diagnostics may or may not be the cause of patients' lower back pain. Diagnostic blocks often precede further interventions in order to confirm or dispute a source of the lower back pain. Chronic lower back pain caused by painful intervertebral disc should be treated using biacuplasty. If the pain of the lumbar spine is caused by facet joints or sacroiliac joint, patient can be offered neuroablative procedures using radiofrequency. In cases where the low back and radicular pain are caused by the central and foraminal spinal stenosis patients are advised epidural steroid injections, unless claudications are present. Patients suffering from the chronic radicular pain may be treated with various nucleoplasty procedures or they may be offered percutaneous radiofrequent neuromodulation. The purpose of above advised procedures is to relieve patients' pain, allow optimal physical therapy, and improved functional capacity, consequently providing better quality of life.

[Guidelines for interventional and invasive treatment in patients with mechanical back pain]. / Smjernice za intervencijske i invazivne postupke u bolesnika s mehanickom krizoboljom.

Back pain caused primarily by mechanical disorders is the most common type of back pain and it is usually found in young and middle-aged population, i.e. active population. This is why back pain is one of the most important public health problems. Treatment of pain syndrome affecting spine depends on a variety of factors and generally includes conservative and invasive methods. Relative indication for interventional and surgical procedures is long lasting back pain, the symptoms of which, predominantly pain, cause significant problems for the patient on performing everyday activities. Invasive/surgical treatment is applied after minimally 3 months of unsuccessful conservative treatment. Invasive and surgical procedures comprise a wide spectrum of interventions, from interventional and semi-interventional procedures, minimally invasive procedures to extensive, invasive operations that include instrumentation. The choice of intervention is based on clinical findings, duration and severity of pain and other symptoms, as well as on diagnostic reports. Despite recommendations based on the results of clinical studies, individual approach to each patient is the main principle of successful treatment.

[New techniques and prospects in invasive treatment in vertebrology]. / Novosti i perspektive u invazivnom lijecenju u vertebrologiji.

In this article authors outline new techniques and prospects in invasive treatment in vertebrology with emphasis on interventional and semi invasive procedures and minimally invasive surgery for lumbar disc herniation. They describe new approaches in neuroablative procedures for back pain treatment, in spinal fixation and in surgical treatment of scoliosis. Authors also report methods of great expectations which are not yet in use in our clinical practice but are promising like reconstruction of nucleus pulposus by autologous chondrocytes transplantation. New methods in cervical spine surgery are also discussed. The efficacy of each surgical method is pointed out. Apart from being informative, together with the corresponding article on patophysiology, magnetic resonance imaging and conservative treatment, these articles considering recent developments can be used as an aid in decision making when approaching these patients.

Peptide therapy with pentadecapeptide BPC 157 in traumatic nerve injury.

We focused on the healing of rat transected sciatic nerve and improvement made by stable gastric pentadecapeptide BPC 157 (10 microg, 10ng/kg) applied shortly after injury (i) intraperitoneally/intragastrically/locally, at the site of anastomosis, or after (ii) non-anastomozed nerve tubing (7 mm nerve segment resected) directly into the tube. Improvement was shown clinically (autotomy), microscopically/morphometrically and functionally (EMG, one or two months post-injury, walking recovery (sciatic functional index (SFI)) at weekly intervals). BPC 157-rats exhibited faster axonal regeneration: histomorphometrically (improved presentation of neural fascicles, homogeneous regeneration pattern, increased density and size of regenerative fibers, existence of epineural and perineural regeneration, uniform target orientation of regenerative fibers, and higher proportion of neural vs. connective tissue, all fascicles in each nerve showed increased diameter of myelinated fibers, thickness of myelin sheet, number of myelinated fibers per area and myelinated fibers as a percentage of the nerve transected area and the increased blood vessels presentation), electrophysiologically (increased motor action potentials), functionally (improved SFI), the autotomy absent. Thus, BPC 157 markedly improved rat sciatic nerve healing.

Traumatic brain injury in mice and pentadecapeptide BPC 157 effect.

Gastric pentadecapeptide BPC 157 (GEPPPGKPADDAGLV, an anti-ulcer peptide, efficient in inflammatory bowel disease trials (PL 14736), no toxicity reported, improved muscle crush injury. After an induced traumatic brain injury (TBI) in mice by a falling weight, BPC 157 regimens (10.0microg, 10.0ng/kgi.p.) demonstrated a marked attenuation of damage with an improved early outcome and a minimal postponed mortality throughout a 24h post-injury period. Ultimately, the traumatic lesions (subarachnoidal and intraventricular haemorrhage, brain laceration, haemorrhagic laceration) were less intense and consecutive brain edema had considerably improved. Given prophylactically (30 min before TBI) the improved conscious/unconscious/death ratio in TBI-mice was after force impulses of 0.068 Ns, 0.093 Ns, 0.113 Ns, 0.130 Ns, 0.145 Ns, and 0.159 Ns. Counteraction (with a reduction of unconsciousness, lower mortality) with both microg- and ng-regimens included the force impulses of 0.068-0.145 Ns. A higher regimen presented effectiveness also against the maximal force impulse (0.159 Ns). Furthermore, BPC 157 application immediately prior to injury was beneficial in mice subjected to force impulses of 0.093 Ns-TBI. For a more severe force impulse (0.130 Ns, 0.145 Ns, or 0159 Ns), the time-relation to improve the conscious/unconscious/death ratio was: 5 min (0.130 Ns-TBI), 20 min (0.145 Ns-TBI) or 30 min (0.159 Ns-TBI).

Possible prognostic significance of p53 and Ki 67 in inverted sinonasal papilloma.

The aim of this study is to test the possible prognostic significance of p53 and Ki67 expression in inverted papilloma of the lateral nasal wall and adjacent sinuses regarding their malignant potential and recurrence. 49 biopsies of the lateral nasal wall and adjacent sinuses obtained from 41 patients from three hospitals were investigated. Immunohistochemically demonstrated p53 and Ki67 expression was measured and statistically evaluated. p53 immunoreactivity was demonstrated in most of papillomas with carcinomas but only in two benign papillomas, while Ki67 demonstrated stronger immunoreactivity in carcinomas and surrounding epithelium. Immunohistochemical staining of inverted sinonasal papillomas for p53 and Ki67 can give useful information concerning the existence of synchronous carcinoma and, in case of high Ki67, a hint toward possible recurrence.

[Treatment of osteoporotic fractures of the spine]. / Mogucnosti lijecenja osteoporotskih prijeloma kraljeznice.

Osteoporosis is a skeletal disease characterized by reduced bone strength due to the loss of bone mass, structural disproportion and reduced calcium level as well as changes in the bone structure caused by trabecular deficit. The loss of bone mass in the vertebrae leads to an increased risk of fractures. Osteoporotic vertebral fractures are two to three times more frequent than hip fractures, especially in women. Osteoporotic compressive fractures of the spine mostly occur in the middle and lower thoracic spine and in the thoracolumbar spine. Common consequences of fractures include long-term pains, decrease in the height and spinal deformity, which significantly correlate with disorders in the gait and mobility, reduced pulmonary function, loss of appetite and in the end increased mortality. Clinical picture in acute conditions manifests itself as a sudden pain in the back following minimal trauma but also often without any trauma at all. Diagnostics is based on a detailed medical history and a thorough clinical examination. Radiological diagnosis relies on standard x-rays. In case of clinical indication (differential diagnosis of tumor or spondylitis, neurological weakness) CT and/or MR imaging is performed. The approach to the treatment of osteoporotic fractures of the spine is individual and is based upon the medical history, localization of symptoms and correlation of the clinical examination with the radiological finding. Open reduction, decompression and fixation are indicated in patients with posttraumatic stenosis and neurological weakness. Current operative methods include vertebroplasty and kyphoplasty, which significantly reduce pain and improve mobility of the patient and consequently, the quality of life.

Differential sensitivity to 4-hydroxynonenal for normal and malignant mesenchymal cells.

Normal and malignant cells of various origin differ in their sensitivity to oxidative stress. Therefore, we used normal and malignant mesenchymal cells--human osteosarcoma cells (HOS and 143B), human fibroblasts (WI38) and two primary cultures of normal human osteoblasts to test sensitivity to reactive aldehyde 4-hydroxynonenal (HNE), known as a second messenger of free radicals and a signaling molecule. Upon HNE-treatment, decrease in cell viability (by Trypan-blue), apoptosis induction (by TiterTACS TUNEL assay), HNE-protein binding (by HNE-His ELISA) were higher in malignant than in normal cells, while glutathione content was higher in normal cells. These results indicate that HNE affects the growth of malignant mesenchymal cells more than normal and that this effect was mainly related to lower glutathione concentration and higher binding of HNE to the cellular proteins. We thus assume that HNE and GSH homeostasis play an important role in the growth regulation of normal and malignant mesenchymal cells.

Pentadecapeptide BPC 157 attenuates chronic amphetamine-induced behavior disturbances.

AIM: To investigate the effect of pentadecapeptide BPC 157 on chronic exposure to amphetamine in rats, particularly the changes commonly referred in chronic amphetamine studies as tolerance (lesser grade of stereotyped behavior, without increased excitability) and reverse tolerance (ie, prominent stereotyped behavior and heightened startle response upon late amphetamine challenges). METHODS: After initial application (initial single dose-regimen), amphetamine (10 mg/kg,ip) was given once daily till d 5 (continuous administration-regimen), and thereafter on d 8, 16, and 46 (intermittent administration regimen). Fo r stereotyped behavior and heightened startle response the observation period was 120 min after amphetamine application, and each animal was observed for 10 s in 5 min intervals. Pentadecapeptide BPC 157 (10 microg/kg or 10 ng/k g, ip) or saline (5.0 mL/kg, ip) were given only at the beginning of the experiment, simultaneously with the initial dose of amphetamine. RESULTS: In relation to applied initial-single/continuous/intermittent amphetamine applications regimen, the control amphetamine rats throughout the experiment showed the changes in stereotyped behavior and heightened startle response, increment or decrement, commonly explained in chronic amphetamine studies as tolerance and reverse tolerance. After t he initial application of the amphetamine, the higher BPC 157 dosage apparently attenuated the stereotyped behavior, while the lower dosage of BPC 157 did not reach a statistical significance. Considering the forthcoming amphetamine challenges, in the rats initially treated with pentadecapeptide BPC 157, either 10 microg- or 10 ng-dose, at the time of the first application of amphetamine, the stereotyped behavior remains to be attenuated after all additional amphetamine challenges (on d 2-5, 8, 16, and 46). This attenuation was not limited to stereotyped behavior only. After the initial application of the amphetamine the heighten ed startle response was also apparently mitigated in rats receiving the BPC 157 dosage, either higher or lower. Later, confronted with the forthcoming amphetamine challenges, they showed apparently less abnormal excitability at all tested points. CONCLUSION: In summary, gastric pentadecapeptide BPC 157 (ie, both microg- and ng-BPC 157 regimens) attenuated chronic amphetamine disturbances. This effect was present throughout the observation period at a statistically significant level. Therefore, it seems that this gastric pentadecapeptide BPC 157 has a modulatory effect on dopamine system, and it could be used in chronic amphetamine disturbances.