Integrated Chemical and Biological Evaluation of Linden Honeydew Honey from Bosnia and Herzegovina: Composition and Cellular Effects

Abstract

Honeydew honey (HH) is a distinctive type of honey known for its dark colour, high mineral and polyphenol content, and pronounced biological activity. This study continues previous research on beech and chestnut honeydew honeys by presenting a comprehensive analysis of linden honeydew honey (LHH) from Bosnia and Herzegovina-a variety that, until now, has not been characterised in detail. Physicochemical parameters confirmed its classification as HH, with high electrical conductivity (1.21 mS/cm) and low moisture (15.1%). GC-MS analysis revealed a unique volatile profile dominated by α-terpinolene (17.4%), distinguishing LHH from other HH types. The sample exhibited high total phenolic content (816.38 mg GAE/kg) and moderate antioxidant capacity (1.11 mmol TE/kg). Antimicrobial testing demonstrated strong activity against Staphylococcus aureus and Methicillin-resistant Staphylococcus aureus (MRSA), with lower efficacy against Gram-negative bacteria. No cytotoxic effects were observed in HaCaT keratinocytes at concentrations up to 60 mg/mL, and wound healing assays showed improved scratch closure reaching approximately 30% after 24 h and 41% after 48 h compared to the control. These results indicate that LHH possesses promising bioactive properties and potential for dermatological application. Further studies with broader sample sets are needed to explore variability and confirm the therapeutic relevance of LHH in comparison to other honeydew types.

Keywords: HaCaT cells; Tilia spp.; antimicrobial activity; antioxidant activity; cytotoxicity; linden honeydew honey; phenolic profile; physicochemical properties; volatile compounds; wound healing.

Figure 1

Map of Bosnia and Herzegovina showing the location where linden honeydew honey (LHH) sample was collected.

Figure 2

Pollen analysis characteristics of the honeydew honey sample.

Figure 3

Inhibition zones (mm) of linden honeydew honey (LHH) sample against different bacteria. Data are presented as mean ± SD (n = 3).

Figure 4

Cell viability of the HaCaT keratinocytes treated with the linden honeydew honey (LHH), assessed using the XTT assay. The percentage of cell viability was measured for different concentrations of honey (expressed in mg/mL of culture medium; range: 0–60 mg/mL). Data are presented as mean ± SD (n = 3).

Figure 5

Influence of the linden honeydew honey (LHH) on the in vitro wound-healing rate 24 h and 48 h period after wounding. The wound-healing rate was expressed as the percentage of scratch closure relative to the initial scratch area. The model wounds were treated with 0.1% honey, and serum-free DMEM served as the control. Data are expressed as the mean ± SD (n = 4). The asterisk (*) represents statistical significance obtained with a two-tailed Mann–Whitney test between the honeydew honey sample and non-template control (NTC) at two time points and significance level p = 0.05.

Figure 6

Representative micrographs of untreated HaCaT cell model wounds (control) and model wounds treated with LHH for 24 and 48 h.