Pharmacologic or genetic targeting of peripheral nerves prevents peri-articular traumatic heterotopic ossification.

Heterotopic ossification (HO) is a pathological process that commonly arises following severe polytrauma, characterized by the anomalous differentiation of mesenchymal progenitor cells and resulting in the formation of ectopic bone in non-skeletal tissues. This abnormal bone growth contributes to pain and reduced mobility, especially when adjacent to a joint. Our prior observations suggested an essential role of NGF (Nerve Growth Factor)-responsive TrkA (Tropomyosin Receptor Kinase A)-expressing peripheral nerves in regulating abnormal osteochondral differentiation following tendon injury. Here, we utilized a recently developed mouse model of hip arthroplasty-induced HO to further validate the role of peripheral nerve regulation of traumatic HO. Nerve ingrowth was either modulated using a knockin transgenic animals with point mutation in TrkA, or local treatment with an FDA-approved formulation of long acting Bupivacaine which prevents peripheral nerve growth. Results demonstrate exuberant sensory and sympathetic nerve growth within the peri-articular HO site, and that both methods to reduce local innervation significantly reduced heterotopic bone formation. TrkA inhibition led to a 34% reduction in bone volume, while bupivacaine treatment resulted in a 50% decrease. Mechanistically, alterations in TGFß and FGF signaling activation accompanied both methods of local denervation, and a shift in macrophages from M1 to M2 phenotypes was observed. In sum, these studies reinforce the observations that peripheral nerves play a role in the etiopathogenesis of HO, and that targeting local nerves represents a potential therapeutic approach for disease prevention.

Integrated transcriptomics of human blood vessels defines a spatially controlled niche for early mesenchymal progenitor cells.

Human blood vessel walls show concentric layers, with the outermost tunica adventitia harboring mesenchymal progenitor cells. These progenitor cells maintain vessel homeostasis and provide a robust cell source for cell-based therapies. However, human adventitial stem cell niche has not been studied in detail. Here, using spatial and single-cell transcriptomics, we characterized the phenotype, potential, and microanatomic distribution of human perivascular progenitors. Initially, spatial transcriptomics identified heterogeneity between perivascular layers of arteries and veins and delineated the tunica adventitia into inner and outer layers. From this spatial atlas, we inferred a hierarchy of mesenchymal progenitors dictated by a more primitive cell with a high surface expression of CD201 (PROCR). When isolated from humans and mice, CD201Low expression typified a mesodermal committed subset with higher osteogenesis and less proliferation than CD201High cells, with a downstream effect on canonical Wnt signaling through DACT2. CD201Low cells also displayed high translational potential for bone tissue generation.

TrkA + sensory neurons regulate osteosarcoma proliferation and vascularization to promote disease progression.

Bone pain is a presenting feature of bone cancers such as osteosarcoma (OS), relayed by skeletal-innervating peripheral afferent neurons. Potential functions of tumor-associated sensory neurons in bone cancers beyond pain sensation are unknown. To uncover neural regulatory functions, a chemical-genetic approach in mice with a knock-in allele for TrkA was used to functionally perturb sensory nerve innervation during OS growth and disease progression. TrkA inhibition in transgenic mice led to significant reductions in sarcoma-associated sensory innervation and vascularization, tumor growth and metastasis, and prolonged overall survival. Single-cell transcriptomics revealed that sarcoma denervation was associated with phenotypic alterations in both OS tumor cells and cells within the tumor microenvironment, and with reduced calcitonin gene-related peptide (CGRP) and vascular endothelial growth factor (VEGF) signaling. Multimodal and multi-omics analyses of human OS bone samples and human dorsal root ganglia neurons further implicated peripheral innervation and neurotrophin signaling in OS tumor biology. In order to curb tumor-associated axonal ingrowth, we next leveraged FDA-approved bupivacaine liposomes leading to significant reductions in sarcoma growth, vascularity, as well as alleviation of pain. In sum, TrkA-expressing peripheral neurons positively regulate key aspects of OS progression and sensory neural inhibition appears to disrupt calcitonin receptor signaling (CALCR) and VEGF signaling within the sarcoma microenvironment leading to significantly reduced tumor growth and improved survival. These data suggest that interventions to prevent pathological innervation of osteosarcoma represent a novel adjunctive therapy to improve clinical outcomes and survival.

Requirement of Pdgfrα+ cells for calvarial bone repair.

Platelet-derived growth factor receptor α (PDGFRα) is often considered as a general marker of mesenchymal cells and fibroblasts, but also shows expression in a portion of osteoprogenitor cells. Within the skeleton, Pdgfrα+ mesenchymal cells have been identified in bone marrow and periosteum of long bones, where they play a crucial role in participating in fracture repair. A similar examination of Pdgfrα+ cells in calvarial bone healing has not been examined. Here, we utilize Pdgfrα-CreERTM;mT/mG reporter animals to examine the contribution of Pdgfrα+ mesenchymal cells to calvarial bone repair through histology and single-cell RNA sequencing (scRNA-Seq). Results showed that Pdgfrα+ mesenchymal cells are present in several cell clusters by scRNA-Seq, and by histology a dramatic increase in Pdgfrα+ cells populated the defect site at early timepoints to give rise to healed bone tissue overtime. Notably, diphtheria toxin-mediated ablation of Pdgfrα reporter+ cells resulted in significantly impaired calvarial bone healing. Our findings suggest that Pdgfrα-expressing cells within the calvarial niche play a critical role in the process of calvarial bone repair.

Mapping Somatosensory Afferent Circuitry to Bone Identifies Neurotrophic Signals Required for Fracture Healing.

The profound pain accompanying bone fracture is mediated by somatosensory neurons, which also appear to be required to initiate bone regeneration following fracture. Surprisingly, the precise neuroanatomical circuitry mediating skeletal nociception and regeneration remains incompletely understood. Here, we characterized somatosensory dorsal root ganglia (DRG) afferent neurons innervating murine long bones before and after experimental long bone fracture in mice. Retrograde labeling of DRG neurons by an adeno-associated virus with peripheral nerve tropism showed AAV-tdT signal. Single cell transcriptomic profiling of 6,648 DRG neurons showed highest labeling across CGRP+ neuron clusters (6.9-17.2%) belonging to unmyelinated C fibers, thinly myelinated Aδ fibers and Aß-Field LTMR (9.2%). Gene expression profiles of retrograde labeled DRG neurons over multiple timepoints following experimental stress fracture revealed dynamic changes in gene expression corresponding to the acute inflammatory ( S100a8 , S100a9 ) and mechanical force ( Piezo2 ). Reparative phase after fracture included morphogens such as Tgfb1, Fgf9 and Fgf18 . Two methods to surgically or genetically denervate fractured bones were used in combination with scRNA-seq to implicate defective mesenchymal cell proliferation and osteodifferentiation as underlying the poor bone repair capacity in the presence of attenuated innervation. Finally, multi-tissue scRNA-seq and interactome analyses implicated neuron-derived FGF9 as a potent regulator of fracture repair, a finding compatible with in vitro assessments of neuron-to-skeletal mesenchyme interactions.

The Mohawk homeobox gene represents a marker and osteo-inhibitory factor in calvarial suture osteoprogenitor cells.

The regeneration of the mammalian skeleton's craniofacial bones necessitates the action of intrinsic and extrinsic inductive factors from multiple cell types, which function hierarchically and temporally to control the differentiation of osteogenic progenitors. Single-cell transcriptomics of developing mouse calvarial suture recently identified a suture mesenchymal progenitor population with previously unappreciated tendon- or ligament-associated gene expression profile. Here, we developed a Mohawk homeobox (MkxCG; R26RtdT) reporter mouse and demonstrated that this reporter identifies an adult calvarial suture resident cell population that gives rise to calvarial osteoblasts and osteocytes during homeostatic conditions. Single-cell RNA sequencing (scRNA-Seq) data reveal that Mkx+ suture cells display a progenitor-like phenotype with expression of teno-ligamentous genes. Bone injury with Mkx+ cell ablation showed delayed bone healing. Remarkably, Mkx gene played a critical role as an osteo-inhibitory factor in calvarial suture cells, as knockdown or knockout resulted in increased osteogenic differentiation. Localized deletion of Mkx in vivo also resulted in robustly increased calvarial defect repair. We further showed that mechanical stretch dynamically regulates Mkx expression, in turn regulating calvarial cell osteogenesis. Together, we define Mkx+ cells within the suture mesenchyme as a progenitor population for adult craniofacial bone repair, and Mkx acts as a mechanoresponsive gene to prevent osteogenic differentiation within the stem cell niche.

Neuronal regulation of bone and tendon injury repair: a focused review.

Beyond the sensation of pain, peripheral nerves have been shown to play crucial roles in tissue regeneration and repair. As a highly innervated organ, bone can recover from injury without scar formation, making it an interesting model in which to study the role of nerves in tissue regeneration. As a comparison, tendon is a musculoskeletal tissue that is hypo-innervated, with repair often resulting in scar formation. Here, we reviewed the significance of innervation in 3 stages of injury repair (inflammatory, reparative, and remodeling) in 2 commonly injured musculoskeletal tissues: bone and tendon. Based on this focused review, we conclude that peripheral innervation is essential for phases of proper bone and tendon repair, and that nerves may dynamically regulate the repair process through interactions with the injury microenvironment via a variety of neuropeptides or neurotransmitters. A deeper understanding of neuronal regulation of musculoskeletal repair, and the crosstalk between nerves and the musculoskeletal system, will enable the development of future therapies for tissue healing.

Accumulating evidence has shown that, across organs systems, peripheral nerves regulate the process of tissue repair and regeneration. This is particularly relevant in the context of musculoskeletal injuries such as those affecting the bone and tendon. The question then arises: what is the function of peripheral innervation in the repair of bone and tendon injuries? This review offers an in-depth look at the ways in which nerves regulate the healing of bone and tendon injuries at various stages of recovery. A deeper comprehension of the influence of nerves on the repair of these tissues could pave the way for the development of future therapeutic strategies for tissue healing.

Biogenic Waste from Two Varieties of Plantain in Ghana Contain Pectin with Potential Binding Properties in Conventional Tablets.

Pharmaceutical formulations have traditionally relied on plants and their derivatives for various APIs and excipients. In Ghana, the widespread utilization of plantains, irrespective of their ripeness, generates significant waste at every stage of processing, posing disposal issues. Fascinatingly, these wastes, often discarded, possess significant economic potential and can be recycled into valuable raw materials or products. Pectin, a polysaccharide that occurs naturally, has seen a surge in interest in recent times. It has found widespread use in the pharmaceutical sector, particularly as a binding agent in tablet formulations. This study aimed to evaluate pectin from two popular plantain varieties, Apem (M) and Apantu (T) at different ripening stages, for pharmaceutical use as a binding agent in immediate-release tablets. The ripening stages selected were the matured-green (G), half-ripe (H), and full-ripe (R). Acid (D) and alkaline (L) mediums of extraction were employed for each ripening stage for both varieties. Wet granulation method was used to prepare the granules using paracetamol as a model drug, and their flow properties were subsequently assessed. Postcompression tests including, hardness, friability, weight uniformity, disintegration, assay, and in vitro dissolution were also assessed. Granules from all formulation batches had good flow properties indicated by their angle of repose (14.93 ± 1.41-21.80 ± 1.41), Hausner ratio (0.96 ± 0.27-1.22 ± 0.02), and compressibility (%) (7.69 ± 0.002-20.51 ± 0.002). All the tablets passed the uniformity of weight with none deviating by ±5%. The hardness of all the formulated tablets ranged between 3.96 ± 0.32 and 13.21 ± 0.36, while the friability for all tablets was below 1%. The drug content was between 100.1 ± 0.23% and 103.4 ± 0.01%. Tablets formulated with pectin as a binding agent at concentrations of 10% w/v and 15% w/v successfully met the disintegration test criteria for immediate release tablets. However, those prepared with a concentration of 20% w/v (MGL, MHD, MHL, MRD, MRL, TGL, THD, THL, and TRL) did not pass the disintegration test. Consequently, all batches of tablets successfully met the dissolution test requirement (Diss, Q > 75%), except for the batches that did not pass the disintegration test (Diss, Q < 75%). Ultimately, pectins extracted from the peels of Apem and Apantu at different ripening stages using acid and alkaline extraction can be commercially exploited as pharmaceutical binders at varying concentrations in immediate-release tablets.

Isolation of Perivascular Mesenchymal Progenitor Cells from Human Adipose Tissue by Flow Cytometry.

Perivascular cells represent an in vivo counterpart of mesenchymal stromal/stem cells that populate the outer layer of blood vessels. Pericytes in capillaries and microvessels and adventitial cells of large arteries and veins give rise to stem/progenitor cells when isolated and cultured in vitro. These cells have been considered candidate cell types for cell therapy. Adipose tissue, being highly vascularized, dispensable, and easily accessed, is a viable option to obtain perivascular cells for use in research and in clinical trials. Here, we describe our established protocol to extract perivascular cells from human fat through fluorescence-activated cell sorting, which allows for the isolation of defined populations of progenitor cells with high reproducibility.

Pharmaceutical Assessment of the Impact of the Method of Extraction on the Suitability of Pectin from Plantain (Musa paradisiaca) Peels as a Suspending Agent in Oral Liquid Formulations.

Natural polymers such as pectin have gained increased utilization in pharmaceutical and biotechnology sectors because they are affordable, easily accessible, nontoxic, and chemically modifiable, with the potential to be biodegradable and biocompatible. Musa paradisiaca (plantain) peels make up 30-40% of the overall weight of the fruit. The extraction of pectin from these residues can therefore be viewed as a possible waste of wealth. This study, therefore, focused on evaluating the suspending properties of pectin obtained from Musa paradisiaca (plantain) peels (through acid and alkaline extraction) and presented an alternative suspending agent in the pharmaceutical formulation of suspensions. The unripe peels of Musa paradisiaca were acquired and authenticated at the Department of Pharmacognosy, Faculty of Pharmacy and Pharmaceutical Sciences, Kwame Nkrumah University of Science and Technology (KNUST), Kumasi, Ghana. Pectin was extracted from the peels using both acid and alkaline extraction processes, respectively, characterized, and evaluated for its phytochemical properties. Different concentrations of the acid and alkaline pectin extracts were employed as a suspending agent in paracetamol suspensions, using acacia gum as a standard. The pectin yields obtained were 4.88% and 7.61% for the acid and alkaline extraction processes, respectively, while phytochemical screening revealed the presence of glycosides, tannins, saponins, and phenols in both extracts. The alkaline pectin extract recorded higher equivalent weight, degree of esterification, ash content, and crude content than the acid pectin extract, while FTIR identified similar functional groups in both acid and alkaline pectin extracts. The test suspensions reported significant differences (P < 0.05) in flow rates, ease of redispersion, sedimentation volumes, and rates compared with acacia gum. Moreover, when the acid and alkaline pectin extracts were compared, significant differences (P < 0.05) were observed in sedimentation rates and sedimentation volumes, suggesting that the extraction method may affect suspending properties. Ultimately, the alkaline pectin extract had better suspending properties than the acid pectin extract; however, they both can be used as an alternative to acacia gum as a suspending agent.

ZIC1 Dictates Osteogenesis Versus Adipogenesis in Human Mesenchymal Progenitor Cells Via a Hedgehog Dependent Mechanism.

Numerous intrinsic factors regulate mesenchymal progenitor commitment to a specific cell fate, such as osteogenic or adipogenic lineages. Identification and modulation of novel intrinsic regulatory factors represent an opportunity to harness the regenerative potential of mesenchymal progenitors. In the present study, the transcription factor (TF) ZIC1 was identified to be differentially expressed among adipose compared with skeletal-derived mesenchymal progenitor cells. We observed that ZIC1 overexpression in human mesenchymal progenitors promotes osteogenesis and prevents adipogenesis. ZIC1 knockdown demonstrated the converse effects on cell differentiation. ZIC1 misexpression was associated with altered Hedgehog signaling, and the Hedgehog antagonist cyclopamine reversed the osteo/adipogenic differentiation alterations associated with ZIC1 overexpression. Finally, human mesenchymal progenitor cells with or without ZIC1 overexpression were implanted in an ossicle assay in NOD-SCID gamma mice. ZIC1 overexpression led to significantly increased ossicle formation in comparison to the control, as assessed by radiographic and histologic measures. Together, these data suggest that ZIC1 represents a TF at the center of osteo/adipogenic cell fate determinations-findings that have relevance in the fields of stem cell biology and therapeutic regenerative medicine.

Assessment of the phytochemical composition and antimicrobial properties of Tapinanthus bangwensis leaves hosted by the branches of Persea americana.

BACKGROUND: Medicinal plants represent a valuable source for new effective and safe antimicrobial drugs making them an alternative therapy. Existing antimicrobial agents are costly and mostly associated with possible side effects. The aim of the present study therefore, was to assess the antimicrobial property and phytochemical composition of hydroethanolic extract of Tapinanthus bangwensis leaves and its fractions. METHOD: T. bangwensis leaves (harvested from its host plant, Persea americana) was extracted by cold maceration with 70% ethanol and further fractionated with different organic solvents using the solvent partitioning method to obtain the crude extract, petroleum ether, chloroform, ethyl acetate and the resulting aqueous fractions. The phytochemical constituents of the extracts were screened and quantified. Also, the TLC of the extracts were analyzed to serve as a fingerprint. Using the agar diffusion and broth dilution methods, the antimicrobial properties of the extracts were assessed. RESULTS: The study showed that the hydroethanolic (70%) crude extract of T. bangwensis leaves and its fractions contain phenolic compounds, flavonoids, saponins, phytosterols and reducing sugars. The phytoconstituents were well extracted into the ethyl acetate fraction than the other fractions evidenced in the high levels (p < 0.0001) of saponins (66.47 ± 1.72% w/w), phenolic compounds (77.75 ± 1.06 mg/100 mg GAE) and flavonoids (44.34 ± 0.06 mg/100 mg QE) contents. From the antimicrobial studies, all the microorganisms tested exhibited varying degrees of susceptibility to the extracts with MIC values between 0.78 to 12.5 mg/mL. The crude extract of T. bangwensis leaves, its ethyl acetate and chloroform fractions also exhibited lethal antimicrobial activity with MLC between 6.25 to 50 mg/mL. CONCLUSION: The crude extract of T. bangwensis leaves and its fractions demonstrated antimicrobial properties against Escherichia coli, Staphylococcus aureus, Staphylococcus saprophyticus and Candida albicans, thereby representing a potential source of natural antimicrobial agent. Further study is required to identify and isolate antimicrobial compounds from the plant for the development of the natural bioactive antimicrobial agents.

"A Review of Parquetina nigrescens (Afzel.) Bullock, A Plant for Traditional Medicine: Phytochemical and Pharmacological Properties".

The development of herbal medicines as a remedy for several illnesses has been recognized and accepted worldwide by health experts. Parquetina nigrescens is a perennial evergreen woody climber from the Apocynaceae family, widely used in Africa for the treatment of many diseases. The current study is intended to review and put together information available on this ethno-medicinal plant, which will improve scientific knowledge about the plant and also identify research areas that need to be investigated further. The information related to the plant was obtained using scientific databases such as Google scholar, WebMD, Wiley, Science direct, Cochrane database, student thesis, PubMed, and Scopus to obtain relevant literature regarding the botanical descriptions and distribution of the plant, traditional uses, phytochemicals, active compounds isolated from the plant, and pharmacological properties of P. nigrescens. Several traditional uses for different parts of the plant (leaves, stem bark, roots, leaf sap, flowers, and latex) have been presented. A review of the phytochemical composition of different plant parts revealed the presence of reducing sugars, flavonoids, tannins, alkaloids, cardiac glycosides, steroids, coumarin, anthraquinones, terpenoids, and saponins. Many studies also highlighted pharmacological activities related to P. nigrescens, including antianemic and haematological activity, antidiabetic, anti-inflammatory, antipyretic, analgesic, antiasthmatic, antimicrobial, insecticidal, neurotoxic, cytotoxic, antityphoid, antipolycystic ovarian syndrome activity, antilipidemic, and memory-enhancing activity. It is recommended that further in-vivo and clinical studies be conducted on the plant for the development of novel drugs.

Bridelia ferruginea Benth.; An ethnomedicinal, phytochemical, pharmacological and toxicological review.

Ethnopharmacological relevance: Bridelia ferruginea belonging to the family Euphorbiaceae, identified as an important commonly growing shrub, is used in traditional medicine for managing arthritis, dysentery, constipation, chronic diabetes, skin diseases, bladder and intestinal disorders, oral infections, thrush, bites and as an arrow poison antidote. This review aims at providing information on the traditional medicinal uses, pharmacological activities, phytochemistry and toxicity studies of Bridelia ferruginea to bridge the gap between traditional medicinal uses and preclinical studies on B. ferruginea and subsequently lead to the development of valued added medicines from B. ferruginea. Materials and methods: Data in this review were compiled using databases such as Google Scholar, Science Direct, Scopus, PubMed, Springer link, Elsevier and Taylor and Francis, articles from peer reviewed journals and other grey literature (short notes, book chapters, short communications) to access all the relevant information available on B. ferruginea. Results: B. ferruginea contains different phytochemicals including flavonoids, phenolics, phytosterols, triterpenes, saponins, alkaloids and cardiac glycosides. Gallocatechin-(4'-O-7)-epigallocatechin, 3,5-dicaffeoylquinic acid, 1,3,4,5-tetracaffeoylquinic acid and some derivatives of 3-methoxyflavone, such as quercetin-3-methyl ether, quercetin 3-,7,3',4'-tetramethyl ether, myricetin 3',4',5'-trimethyl ether, myricetin 3,3',4',5'-tetramethyl ether, myricetin and quercetin 3-O-glucoside specific flavonoids and biflavonoids like apigenin, kaempferol and glycosides of both have been isolated and further characterized from B. ferruginea. B. ferruginea has several pharmacologically beneficial properties including anti-inflammatory, anti-diabetic, antioxidant, antimicrobial, anti-infective, antipyretic, analgesic, diuretic and natriuretic activities. Conclusion: The wide distribution, traditional medicinal uses and wealth of phytochemicals present in B. ferruginea suggests that the plant can be useful in lead compound discovery. Although B. ferruginea has been widely studied, further studies on the mechanism of action, bioavailability, pharmacokinetics, toxicity and side effects in humans need to be investigated.

Development and In Vitro Evaluation of Oral Capsules from Antiaris: A Convenient Substitute for Peripheral Neuropathy.

Antiaris is a monoherbal decoction produced by the Centre for Plant Medicine Research (CPMR), Mampong-Akuapem, Ghana. It is prepared from the stem bark of Antiaris africana Engl. (Moraceae), prescribed, and dispensed to patients for the management of nervous disorders. This current formulation presents notable challenges in patients' adherence to treatment regimen due to its bulkiness and bitterness. These challenges have resulted in a decrease in therapeutic outcome. This study sought to transform Antiaris into oral capsules to mask its bitter taste and reduce bulkiness of the product to improve patients' convenience. In this study, four (4) conventional release capsule formulations were successfully prepared from the decoction via wet granulation using corn starch, lactose, light magnesium carbonate (LMC), and microcrystalline cellulose (MCC) and labelled A01, A02, A03, and A04 respectively. The drug-excipient compatibility studies on A01, A02, A03, and A04 were investigated using Fourier transform infrared (FTIR) spectroscopy. The flow properties of the granules as well as the quality assessment of the formulations such as dissolution, disintegration, uniformity of weight, and assay tests were evaluated using pharmacopoeial and nonpharmacopoeial methods. Appropriate models were used to investigate the difference factor (f 1 ) and similarity factor (f 2 ) of the dissolution profiles of the formulations and Antiaris. From the study, all formulated granules had excellent flow properties with Carr's index from 7.83 to 9.56%, Hausner's ratio from 1.09 to 1.10, and angle of repose from 25.13 to 27.87°. Drug-excipient compatibility studies demonstrated no interaction between extract and used excipients. All formulations passed the uniformity of weight, disintegration, assay, and dissolution tests. Formulation A02 had the highest dissolution efficiency of 100.12%, while A03 recorded the least value of 97.22% in the 1 h dissolution studies. A comparison of their various dissolution profiles, respectively, to that of its decoction demonstrated their similarity, since, in all comparisons, f 2 < 15 and f 1 > 50. This implies that, any of these four formulations could be a good substitute for Antiaris.

Approaches for the Elimination of Microbial Contaminants from Lippia multiflora Mold. Leaves Intended for Tea Bagging and Evaluation of Formulation.

Elimination of microorganisms from herbal products has been a major concern due to its implicated health risk to consumers. Drying of herbal materials has been employed for centuries to reduce the risk of contamination and spoilage. The present study adopted three drying approaches in an attempt to eliminate microorganisms from Lippia multiflora tea bag formulation. This study also evaluated the tea bags and optimized the extraction procedure. The L. multiflora leaves for tea bagging were air-dried and milled (A), oven-dried and milled (B), and microwaved (the milled air-dried leaves) (C). The moisture contents were determined at 105°C ± 2°C for 2 hours to constant weight. Phytochemical parameters such as phytochemical constituents, total water extractive, and pH were assessed. The microbial safety and quality of the L. multiflora tea bags were evaluated using the British Pharmacopoeia 2019 specifications. The uniformity of the mass of the formulated tea bags was also determined. Extraction from the Lippia tea bags was optimized. The results showed that using the approaches (A, B, and C) adopted for drying and processing, the moisture contents of the formulated tea bags were in the range of 9.75-10.71% w/w. All the formulated tea bags contained reducing sugars, phenolic compounds, polyuronides, flavonoids, anthracenosides, alkaloids, saponins, and phytosterols. The pH range of the formulations was 7.11-7.54, whereas the total water extractive values were in the range of 19.12-20.41% w/w. The one-way analysis of variance demonstrated no significant difference in the data obtained from the results from A, B, and C. The formulation from A was found to be unsafe for consumption due to unacceptable microbial contamination limits. Microbial load of the formulations from B and C were within the BP specifications. All the batches of the formulations passed the uniformity of mass test. An optimized extraction procedure was obtained when one tea bag was extracted in 250 mL of hot water within the specified time. L. multiflora leaves meant for tea bagging should be oven-dried or microwaved before tea bagging for safe consumption.

A review on anti-peptic ulcer activities of medicinal plants used in the formulation of Enterica, Dyspepsia and NPK 500 capsules.

Peptic ulcer disease affects many people globally. With the increasing resistance to some orthodox antibiotics such as Clarithromycin and Metronidazole, it is important that new acceptable, safer and effective therapies are developed to manage this disease. Various herbal medicines have been used traditionally for the remedy of peptic ulcer disease (PUD), however scientific information with regards to their anti-peptic ulcer both in-vivo and in-vitro as well as clinical studies supporting their use is still inadequate. The Centre for Plant Medicine Research, (CPMR) Mampong-Akuapem, Ghana manufactures three herbal Products namely Enterica, Dyspepsia and NPK 500 capsules which are currently used for the remedy of PUD as a triple therapy at its out-patient clinic with promising effects. The aim of this review is to gather information from literature on the anti-ulcer properties, pharmacological, phytochemical constituents and related activities of herbal plants used at the CPMR for formulation of the triple herbal therapy. This review may, provide some scientific bases for the use of Enterica, Dyspepsia and NPK 500 capsules in the management of Peptic ulcer at the CPMR out-patient clinic. METHODS: Organization for the review involved the on and/or offline search for information from available literature using electronic data and scientific research information resources such as PubMed, Science Direct and Google scholar. RESULTS: In this review, fifteen ethno-medicinal plants used for the formulation of Enterica, Dyspepsia and NPK capsules have been discussed, presenting the description of the plants, composition and pharmacological activity. INTERPRETATION: Tables with the summary of reviewed medicinal plants with their anti-ulcer models and inference on possible mechanisms of action were drawn up. The mechanism(s) of action of individual plants and products (Enterica, Dyspepsia and NPK 500 capsules) must be further investigated and established experimentally in-vitro in addition to in-vivo pharmacological and clinical activity studies to confirm their use in the remedy of PUD.