Osteopenia presents a standard of bone density below average but not to the same extent as osteoporosis. Bone densitometry T scores ( -1 to -2.5) present it according to the World Health Organization for describing it. Osteopenia can be brought on by various factors, such as inactivity, a lack of calcium, and vitamin D [1]. Genetics has a significant impact on bone mineral density, and osteopenia is typically found in premenopausal Caucasian women with thin frames. Treating calcium and vitamin D deficiency and walking 3 to 5 miles per week is frequently possible to increase bone density in the hip and spine [2].

Genetics has a significant impact on bone mineral density, and osteopenia is typically found in premenopausal Caucasian women with thin frames. Bone density in the hip and spine may be improved by addressing calcium and vitamin D deficiencies and engaging in 3 to 5 miles of weekly walking [3]. Osteocalcin (OC), a byproduct of osteoblasts, accumulates in the extracellular matrix of bone. The level of serum OC, a well-known marker of osteoblast activity, reveals how quickly bones are created. It is primarily produced by osteoblasts and is used as a marker to determine how quickly bones are being replaced [4]. The histomorphometry markers of bone development and the serum OC level are sensitive indicators of bone synthesis. Although its precise role within the bone matrix is still unclear, it is associated with a higher rate of bone turnover and a lower BMD [5].

The capacity to create and release tiny antimicrobial peptides is one of the most crucial elements of innate immunity. Recent research has suggested that antimicrobial peptides (AMPs) are critical immunological variables. Some of these AMPs are human-defensins (HBD). Multipurpose peptides called AMPs serve as the initial line of defense against various infections [6]. Defensins are a class of organic AMPs that have potent antiviral, antibacterial, and antifungal properties. Additionally, defensins are considered multifunctional components [7]. All three human beta-defensins were present in the mineralized bone matrix at the endosteum and osteocytes of the chronically infected mandibular bone. In the cytoplasm of osteocytes, hBD-1, -2, and -3 were also found [8]. All three beta-defensins were expressed in all non-infected bone types, consisting of the controls, albeit to a lesser degree than that seen in the chronically infected mandibular bone [9, 10]. The current in vitro research aimed to examine the levels of human defensin-2 and osteocalcin in menopausal women with osteopenia.

Type of Study

Prospective study.

Code of Ethics

Our publication ethics and publication malpractice statement are mainly based on the Code of Conduct and Best-Practice Guidelines for Journal Editors (Committee on Publication Ethics, 2011).

Inclusion Criteria

• All women between 40-50 years old with T-score -1.0 or more are considered normal.
• T-score between -1.0 to -2.5 is classified as osteopenia.
• T-score -2.5 or less is labeled as "osteoporosis."

Exclusion Criteria

Men, women below 40, and those above 50.

Sampling

The menopausal state was defined by the absence of menses for more than a year in females 50 or older. Ninety (90) postmenopausal women, aged between 50-67 years, were enrolled in this research and divided into two groups: A) fifty (50) women with osteopenia and B) forty (40) serving as healthy controls. Bone mineral density (BMD) was measured using dual-energy x-ray absorptiometry (DEXA) scans for all women attending the Rheumatology and Rehabilitation outpatient clinic at Baghdad Teaching Hospital between July 2022 and September 2022. The World Health Organization (WHO) criteria were used for diagnosis:

• T-score -1.0 or more is normal.
• T-score between -1.0 to -2.5 is osteopenia.
• T-score -2.5 or less is osteoporosis [11].

Serum studies included the measurement of calcium, phosphorus, and alkaline phosphatase using a spectrophotometer, as well as the measurement of serum human-defensin-2, osteocalcin, and vitamin D using enzyme-linked immunosorbent assay (ELISA) with a kit from CUASBIO, Chania.

Patients and healthy controls were excluded if they were smokers and drinkers, had a history of illnesses impacting bone metabolism, including endocrine disorders, gastrointestinal tract illnesses, renal diseases, and hematologic disorders, had rheumatoid arthritis, liver conditions, or were using drugs known to impact bone turnover (Steroid therapy, thyroxine, heparin, barbiturates, phenytoin, and Thiazolidinediones).

Collection of Samples

Approximately 5 ml of venous blood at room temperature was deposited in the tube and centrifuged at \(3 \times 10^3\) rpm for five minutes. The serum was then placed in an Eppendorf tube and stored in a freezer set at -20 \(^\circ\)C. Body mass index was calculated using a specific equation.

Statistical Analysis

SAS was used to handle the data (Statistical Analysis System - version 9.1). The significance of the discrepancies between the means was determined using an impartial t-test. Statistics were considered significant if \(P \leq 0.05\) [12].

HBD2 (pg/ml), ALP(IU/L) and osteocalcin elevated in patients when compared to control samples. Calcium and 25OH Vit. D levels were less in patients than in control.

BMI, or body mass index, was discovered to be significantly different between patients and controls (p 0.01) despite age and menopausal age being similar between the two groups. Although differences in serum levels of calcium, phosphorus, vitamin D, also alkaline phosphatase had seen among illness people (as a sample) , and controls, serum levels of HBD2 also osteocalcin have considerably lower in controls in comparative with patients, see Table 1.

T-score is the best a defining characteristic among patients’ osteopenia and control. Table 2 shows the comparison between patients and control group in T-score and Z-score which show significant difference (p<0.01).

Menopause Age (year) and osteocalcin ng/ml have a substantial but somewhat favorable association, according to the individual correlation results (r= .456, p < .001),and significant medium positive relationship between Menopause Age (year) and Calcium mg/dl , (r= .346, p = .014), as shown in Figures 1 and 2 respectively.

Figure 1: The Correlation Coefficient r between Menopause Age and Calcium Level

Figure 2: The Correlation Coefficient r between Menopause Age and Osteocalcin Level

Our findings show an increase in people beta defensin 2 and osteocalcin standards in menopausal women with osteopenia HBDs and have been found to stimulate bone remodeling and have immunomodulatory protective properties [13]. Osteopenia and osteoporosis are frequent conditions that are linked to an increased risk of fractures, particularly in the aged population. These biochemical indicators provide information on the dynamic state of bone metabolism. Age, gender, and menopausal state all impact the indicators of bone turnover, which are further broken down into markers of bone creation and markers of bone breakdown [14]. Studies qualified thus far propose that human \(\beta\)-defensin 2 is a substantial protein of innate immune reaction that supplies defense about the people organism versus invading of all pathogens [15]. It may also be considered a marker of inflammation standards of beta-defensins have so serious in postmenopausal illness people in some unrest, such as lichen sclerosis, because reduced condensation of hBD-1 and significantly excessed levels of hBD-2 and hBD-3 in females are substantive [16]. Overproduction of protection transposes into raised action of fibroblasts and production of the extracellular matrix, that elevate illnesses advancement. This raises the prospect of curing these illnesses by altering the microbiome’s makeup that causes them [17].

Menopause is associated with a broad alteration in the metabolic features consideration [18, 19]. Menopause causes metabolic and endocrine changes in women, as well as the cessation of fertility, resulting in fat cumulation in the body, the expansion of focal obesity, and a rise in insulin resistance, which leads to MetS. Osteocalcin (OCN), which is produced specifically by osteoblasts and is the most abundant non-collagenous protein in bone, has been shown to inhibit bone formation [20] preceding scientific records the serum osteocalcin concentration had influenced by age, female or male, strain [21, 22], and menopausal case. High bone turnover occurs in women during the menopausal transition, according to the bone turnover marker osteocalcin. Even though postmenopausal women’s serum osteocalcin levels were greater than those of either control or premenopausal individuals [23, 24]. Our study reported a decrease in vitamin D and calcium levels in postmenopausal women; the normal value of vitamin D and Ca intake is the cornerstone of osteoporosis prevention and treatment. Age-related changes in vitamin D and calcium metabolism increase the risk of vitamin D insufficiency and secondary hyperparathyroidism. Although previous studies suggested a role of vitamin D intake in modulating bone loss in perimenopausal [25] women, studies of vitamin D and calcium supplementation have failed to support a significant effect of vitamin D and calcium during early menopause. There is a more apparent benefit in vitamin D and calcium supplementation in older postmenopausal women. Vitamin D intake between 500 and 800 IU daily, with or without calcium supplementation, has increased bone mineral density (BMD) in women with a mean age of approximately 63 years.

Increasing serum HBD 2, osteocalcin levels in postmenopausal women with osteopenia plays an important role in development of osteopenia to osteoporosis.

Acknowledgment to Wasnaa J., Sanan Th., Anfal S. Sameen who participate effectively in this article and Deep appreciation to all the staff at the Baghdad Teaching Hospital in the Medical City. Baghdad, Iraq.

This research paper received no external funding.

The authors declare no conflicts of interest.

All authors contributed equally to this paper. They have all read and approved the final version.

Informed consent was obtained from all participates in the study as needed.

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